Integration of microfluidics and ART a new frontier in gamete and embryo handling

Introduction

ART technology has progressed significantly to treat infertility in both humans and animals. The conventional gamete manipulation techniques as well as fertilization methods and embryo culture systems remain both labor-intensive and poorly standardized and operator-dependent which can negatively affect fertilization success rates and embryo development (Swain et al., 2013) [42] . Microfluidics a discipline uniting physics with engineering and biology and chemistry represents an effective answer to existing problems. Microfluidics manipulates fluids within microliter to picoliter volumes through microscale channels which enables strict control over the physical and chemical microenvironment (Whitesides, 2006) [44] . The precise control enables better nutrient distribution while maintaining efficient waste removal and real-time observation of gametes and embryos under regulated settings (Berthier et al., 2012; Sackmann et al., 2014) [3, 35] . The ART field utilizes microfluidic platforms to perform sperm sorting procedures alongside oocyte handling and IVF processes and embryo culture applications. The systems function as sperm selection tools which evaluate motility as well as rheotaxis and chemotaxis and thermotaxis and surface charge properties that conventional swim-up and density gradient centrifugation fail to assess (Ricci et al., 2009, Jahangiri et al., 2024) [33,20] . Through microfluidics scientists achieve delicate oocyte manipulation while decreasing mechanical stress which enhances both maturation and developmental potential (Lai et al., 2015; Yanez and Camarillo, 2017) [24,47] . The integration of microfluidics with 3D culture systems and organ-on-a-chip platforms enables better replication of in vivo conditions through recent technological developments. These advanced models enable better studies of embryo-maternal interaction while providing cell differentiation in environments that duplicate natural reproductive tissues (Pensabene et al., 2026; Stern-Tal et al., 2020; Gargus et al., 2020) [32,40,14] . This paper examines the fundamental elements of microfluidics together with its connections to ART practices. The discussion explores current applications in sperm and oocyte management and embryo cultivation and the joint implementation with new technologies including 3D matrices and bioprinting. The review examines present obstacles including standardization requirements and biocompatibility issues and clinical workflow integration while providing perspective on in vitro embryo production advances through microfluidic technology.

Basic Principles of Microfluidics

The field of microfluidics integrates microengineering with fluid mechanics and biological applications to manage tiny fluid volumes between picoliters and microliters through microchannel systems (Whitesides, 2006; Ma et al., 2024) [44,28] . The fabrication of these devices through PDMS glass and thermoplastics enables unmatched precision for cell and biofluid and gamete management under tightly controlled conditions. Fluids show laminar flow behavior at the microscale because their Reynolds numbers remain low which causes them to flow in parallel non-mixing layers. The ability to predict diffusion-driven transport exists because of this feature which also eliminates shear stress that occurs in turbulent systems (Stone et al; 2004; Squires and Quake, 2005) [41,39] . The small size of microchannels allows them to provide large surface-to-volume ratios which boost heat transfer rates and chemical exchange rates required to support stable microenvironments for cells and embryos (Berthier et al., 2012; Xiao et al., 2017) [3,46] . Several microscale components that include channels mixers valves and chambers work with external actuation mechanisms like pressure gradients electrokinetics magnetophoresis or temperature fields to achieve fluid control according to Sackmann et al. (2014) [35] and Nosrati et al. (2014) [31] The forces provide non-invasive handling of gametes and embryos through low reagent consumption.Modern microfluidic chips now include builtin sensors which measure temperature and pH and oxygen levels to provide continuous embryo monitoring during in vitro development (Zhang et al., 2021) [50] . The fundamental importance of microfluidics in modern assisted reproductive platforms emerges from its automated precise and repeatable biological workflows (Swain et al., 2013) [42] .

Benefits and Motivation for Miniaturization in Microfluidics

The miniaturization of fluidic environments enables microfluidics to achieve multiple advantages above conventional macroscale systems. The microscale manipulation of fluids delivers improved operational efficiency along with reduced costs and heightened sensitivity and better control that benefits applications such as assisted reproductive technologies (ART). [35,26] .  High Sensitivity and Reproducibility: The small size of microfluidic systems enables accurate control over their microenvironment which leads to better biochemical reaction sensitivity and dependable results (Ma et al., 2024) [28]  Rapid Reaction and Analysis Time:

Key Benefits of Microfluidics

The short reaction time possible in microfluidic systems results from small sample volumes that allow for faster mixing and diffusion and thermal equilibrium (Xiao et al., 2017) [46] .  Enhanced Mass and Heat Transfer: The high surfaceto-volume ratio of microfluidic devices enables efficient transfer of nutrients as well as gases and heat which suits embryo culture requirements (Berthier et al., 2012) [3] [50] .

 Minimized Contamination and Evaporation:

The closed-channel design protects against external contaminants and the low thermal mass allows rapid temperature and pH stabilization (Stone et al., 2004) [41] .  Single-Cell and Single-Sperm Manipulation: Microfluidic systems achieve the separation and examination of individual gametes through their ability to evaluate their motility and surface charge properties and morphological features (Jahangiri et al., 2024) [20] .

Motivation behind Miniaturization in ART

ART applications benefit from miniaturization since it creates environments that closely resemble natural conditions. The human female reproductive system includes small fluid channels that display laminar flow together with specific chemical balance gradients. A microfluidic chip that replicates these conditions enables better physiological handling of gametes and embryos (Xiao et al., 2017) [46] . The minimal mechanical stress in microfluidic systems protects fragile biological cells like oocytes and blastocysts while simultaneously boosting their survival rates and developmental outcomes (Lai et al., 2015; Gargus et al., 2020) [24,14] . Automation becomes feasible through miniaturization which enables essential clinical reproducibility for IVF and sperm sorting in veterinary and human medical contexts (Sackmann et al., 2014; Ferraz et al., 2023) [35,10] . [23,20] . Microfluidics enables biomimetic functional selection which duplicates how sperm naturally choose the oocyte by utilizing cues including fluid flow chemical gradients and temperature signals. The technologies permit selection through motility rheotaxis chemotaxis thermotaxis boundary following and surface charge while keeping DNA integrity intact.

Sperm Selection Using Microfluidics

Motility-Based Microfluidic Selection

The quality of sperm depends heavily on its motility. Microfluidic systems from the past utilized sperm swimming against fluid flow and nutrient-rich areas to select motile sperm passively according to Cho et al., 20 [6] and Gadelha et al. (2019) [13] . These platforms eliminate centrifugation steps, reducing reactive oxygen species (ROS) and DNA damage. Sperm motion consists of linear flagellar beating as well as multiple complex biomechanical interactions which occur between the fluid environment. The properties of sperm motility enable microfluidic systems to extract sperm cells with the best possible motility characteristics (Norsati et al.,2014., Gadelha et al., 2019) [31,13] .

Spartan (Simple Periodic Array for Trapping and isolation):

The innovative microfluidic system SPARTAN utilizes flagellar kinematics to differentiate sperm.

The system contains periodic micropillar arrays that capture sperm with irregular flagellar movements or shapes yet permits normal motile sperm to pass through (Nosrati et al., 2014) [31] . SPARTAN-selected sperm show higher DNA integrity, better morphology, and improved fertilization potential. The method operates without labels or centrifugation agents which makes it appropriate for clinical settings.

Rheotaxis-Based Sperm Selection

Sperm exhibit rheotaxis as a natural trait through their ability to swim opposite to fluid flow when found in oviduct environments (Miki and Clapham, 2013) [29] . The artificial simulation of directional fluid flow in microfluidic devices enables the separation of highly mobile sperm with improved DNA quality and better structural integrity (Kantsler et al., 2014; El Sherry et al., 2023) [22,9] . Modern rheotaxis-based chips use structured channels or corrals that induce flow gradients. The collection zone remains accessible only to sperm which possess strong motility along with rheotactic properties while all other sperm get eliminated by the process (Zaferani et al., 2019) [49] . The devices provide sperm selection with non-invasive capabilities while being label-free and capable of high throughput operations. Microfluidic sperm sorting platforms utilize physiological guidance mechanisms such as rheotaxis, chemotaxis, thermotaxis, and surface chargebased separation to improve functional sperm selection (Figure 1)

Chemotaxis-Based Sperm Selection

The biochemical guidance mechanism chemotaxis enables sperm to detect chemical gradients that come from the oocyte and cumulus cells (Eisenbach et al., 2025) [8] . Microfluidic chips generate gradients to separate the sperm subpopulation which demonstrates maximum sensitivity to these cues and shows optimal fertilization potential within the range of 10-15%. Channels receive attractants such as progesterone and acetylcholine and heparin to create conditions similar to the body's natural environment. Sperm selection based on chemotaxis improves the capacitation process and acrosome reaction and zona binding capacity (Eisenbach et al., 2025) [8]

Thermotaxis-Based Sperm Selection

The oviductal isthmus and ampulla experience a small temperature variation (0.5-2°C) during ovulation. The process of thermotaxis helps sperm cells to migrate toward warmer temperature zones for successful fertilization (Bahat and Eisenbach, 2006) [2] . These microfluidic systems create stable temperature differences across their channels through their design. The sperm selection process using thermotaxis results in better DNA quality and improved fertilization outcomes according to research conducted with thermal microfluidic chips (Li et al., 2014; Wu et., 2024) [27,45] .

Boundary Following

Sperm cells exhibit boundary following as a wallguided behavior by showing alignment and movement along the walls of microchannels (Samuel et al., 2018) [36] . Sperm head-wall interactions along with flagellar dynamics determine the way sperm behave in this context. The fallopian tube conditions are replicated by microfluidic chips containing circular or spiral microchannel arrays to find sperm that are stable without needing centrifugation (Khodamoradi et al., 2021) [23] . Some samples may experience decreased efficiency because of wallrelated clogging or debris accumulation.

Surface Charge (Zeta Potential)

The sperm membrane carries negative charges from sialic acid composition that scientists measure as zeta potential. The zeta potential values of functionally competent sperm differ from those of immature sperm cells or apoptotic sperm cells according to Esfahani et al. (2016) [30] . Sperm sorting occurs through dielectrophoresis and electrostatic interactions which function on microfluidic devices that use electric fields to separate sperm based on their surface charge. The testicular sperm extraction (TESE) process can benefit from these methods to extract non-motile sperm in asthenozoospermia cases (Iranzo et al., 2025) [19] . [24,47] .

a. Microfluidic Denudation and Handling

The removal of cumulus cells becomes crucial because it allows ICSI procedures and embryo evaluation to take place. The combination of micropillar arrays and flowshear interfaces in microfluidic devices enables the efficient removal of cumulus cells through mechanical methods which replaces both enzymatic digestion and pipetting stress (Han et al., 2010) [16] . The mechanical denudation technique protects oocytes from damage and maintains their zona pellucida structure.These platforms also automate the capture and positioning of oocytes which decreases both operator dependence and the chances of contamination (Yanez et al., 2016) [48] .

b. Oocyte Selection and Maturation

Oocyte selection methods in traditional practices depend on manual visual assessment of morphological characteristics. Modern microfluidic systems now integrate optical and mechanical sensors to conduct real-time measurements of oocyte dimensions and zona pellucida thickness as well as cytoplasmic texture and shape (Yanez and Camarillo, 2017) [47] . A microfluidic chamber that includes cumulus or granulosa cells enables the simulation of natural ovarian conditions which improves oocyte maturation (Han et al., 2010) [16] . This method enhances the success of fertilization by improving both meiotic development and cytoplasmic maturation.

c. Microfluidics in Cryopreservation

Osmotic shock together with ice crystal formation represents the main challenges during conventional cryopreservation methods. The microfluidic chip technology enables sequential exposure to cryoprotectants through precise control of flow rates and temperature gradients and CPA gradients (Lai et al., 2015; Guo et al) [24] . These devices enable the reduction of oocyte shrinkage and they enhance post-thaw survival as well as spindle integrity and cleavage rates. Microfluidics-enhanced vitrification through murine and bovine models produced superior results in terms of morphology and membrane integrity and developmental competence (Aydin et al., 2024) [1] .

In Vitro Fertilization, Embryo Culture, and Analysis using Microfluidics 1. Microfluidics in In Vitro Fertilization (IVF)

IVF microfluidic systems duplicate the spatial-temporal dynamics of female reproductive tract to enable controlled gamete interactions. The systems decrease polyspermy occurrences while maintaining sperm-oocyte synchrony and minimizing shear stress that directly affects zygote development (Richards et al., 2024) [34] . IVF chips that consist of microwell arrays enable direct observation of fertilization events and prevent the zygote overcrowding effect. The advanced devices possess the capability to control sperm numbers and concentrations when introducing sperm to each oocyte especially useful during cases of low sperm count (Gurkan et al., 2024) [15] .

Embryo Culture in Microfluidic Systems

Traditional culture systems often fail to mimic the in vivo uterine environment. The microfluidic platforms generate stable microenvironments that replicate all in vivo mechanical forces and pH conditions and nutrient flow patterns. The systems require fewer medium exchanges and sustain optimal embryonic environments through continuous perfusion (Lee et al., 2021; Ren et al., 2015) [26,51] . Embryos cultured in microfluidic systems show improved blastocyst development, enhanced cell number and differentiation, better mitochondrial activity and epigenetic stability (Zhao et al., 2015) [51]

Real-Time Embryo Monitoring and Metabolic Assays

The built-in sensors in microfluidic embryo culture chips provide non-invasive real-time monitoring of embryo viability. The use of electrochemical or optical sensors in these systems enables continuous tracking of glucose, lactate, pH and oxygen tension to evaluate embryo metabolic activity and developmental capacity (Heo et al., 2010; Swain et al., 2013) [17,42] . The analytical microfluidic systems enable the selection of high-quality embryos without the need for biopsy thus supporting safer and more accurate embryo transfer decisions. Microfluidic IVF offers several advantages over conventional IVF systems because it more closely mimics the physiological microenvironment of the female reproductive tract. In microfluidic IVF, the culture medium volume is typically maintained at very low levels (1-5 µL) compared to conventional IVF systems that generally require 100-500 µL of media. This reduced volume enhances the accumulation of embryo-secreted autocrine and paracrine factors while simultaneously decreasing reagent consumption (Heo et al., 2010) [17] . In addition, gamete handling in microfluidic systems occurs within enclosed microchannels and is largely automated, thereby minimizing manual pipetting and centrifugation procedures commonly used in conventional IVF laboratories. As a result, microfluidic platforms reduce mechanical stress, oxidative stress, and DNA damage to gametes and embryos (Swain et al., 2013) [42] . Furthermore, these systems provide controlled fluid dynamics, continuous nutrient exchange, and opportunities for real-time, non-invasive monitoring of embryo development through integrated sensing technologies (Bhatia and Ingber, 2014) [5] . Overall, microfluidic IVF systems more effectively reproduce in vivo reproductive conditions and therefore have substantial potential to improve fertilization outcomes, embryo quality, and developmental competence (Xiao et al., 2017) [46] .

3D Culture Systems for Gametes and Embryo Manipulation

The conventional 2D culture systems restrict cells to flat rigid surfaces which lack physiological significance. In contrast, 3D culture systems allow gametes and embryos to interact in all three dimensions, mimicking the extracellular matrix (ECM) and in vivo reproductive environments more closely (Laronda et al., 2017; Jensen and Teng, 2020) [25,21] . The early stages of embryonic development require precise mechanical cues and spatial orientation and cellular signaling which 2D systems fail to provide adequately (Sittadjody et al., 2017, Hoffman, 2018) [38,18] . Different biomaterials such as type I collagen, gelatin, and alginate hydrogels have been employed for 3D embryo culture. Soft scaffolds made from collagen at a stiffness of 1 kPa support better cleavage rates along with improved blastocyst development and hatching and cell lineage differentiation (Zhao et al., 2015) [51] . The biocompatible and non-cytotoxic alginate hydrogels derived from brown algae allow for embryo elongation and adhesion and morphogenesis and are tunable (Zhao et al., 2015; Zhao et al., 2015) [51,51] . Porous alginate scaffolds also enable coculture with oviductal or endometrial cells to simulate implantation environments (Stern-Tal et al., 2020) [40] . The platforms enhance embryo viability while also improving morphology and trophoblast differentiation which are essential for implantation success and postnatal outcomes. The 3D matrices provide suitable environments for in vitro growth of follicles and maturation of oocytes. Hydrogelbased ovarian follicle encapsulation techniques have been utilized across mice and human and cattle and dog species for ex vivo oocyte production (Gargus et al., 2020) [14] . They offer Controlled stiffness to support antral follicle expansion, improved oxygen and nutrient diffusion, Coculture support with granulosa cells or hormones to mimic follicular fluid conditions. Compared with conventional two-dimensional (2D) culture systems, three-dimensional (3D) culture platforms provide a more physiologically relevant microenvironment for gamete and embryo development. In 2D culture, cells grow on flat rigid surfaces with limited spatial organization and restricted cell-cell interactions, often resulting in altered embryo morphology and suboptimal developmental outcomes. In contrast, 3D culture systems mimic the native extracellular matrix and support dynamic multidirectional cellular interactions, allowing embryos to maintain natural spheroid or tubular morphology. These systems also facilitate implantation-like processes through co-culture approaches and improve embryo viability, trophoblast differentiation, and placental development after transfer (Hoffman, 2018; Jensen and Teng, 2020) [18,21] . Biomaterials such as collagen and alginate hydrogels have been widely used to create supportive 3D reproductive microenvironments that better resemble in vivo conditions (Zhao et al., 2015; Stern-Tal et al., 2020) [51,40] .

Organ-on-a-Chip and Bioprinting in Reproductive Technologies

The technology of Organ-on-a-chip produces microengineered 3D models which reproduce the structure and functional behavior of actual tissues. The technology unites microfluidics with cell culture methods along with sensor systems to generate organ-scale physiological and disease-related behavior (Bhatia and Ingber, 2014) [5] . ART researchers and reproductive biology scientists utilize organ on a chip platform to recreate and investigate ovarian follicle growth, menstrual cycle hormone dynamics, embryo-maternal interactions, implantation and endometrial receptivity (Xiao et al., 2017, Pensabene et al., 2026) [46,32] . Applications of organ-on-a-chip and bioprinting technologies in reproductive biology and ART are summarized in Figure 2.

Key Reproductive Organs Modeled on Chips

 Ovary-on-a-Chip: The device supports the in vitro growth of both primordial and antral follicles through periodic hormone treatments. These chips enable the process of oocyte maturation together with steroidogenesis (Gargus et al., 2020) [14] .  Endometrium-on-a-Chip: A device duplicates either a receptive or non-receptive uterine lining condition.

Stromal and epithelial cell co-culture under hormonal exposure generates implantation models that are realistic (De Bem et al. Stern-Tal et al., 2020) [40]  Embryo-maternal interface: The technology allows researchers to examine blastocyst adhesion as well as hatching and nutrient exchange when applying flow conditions. These platforms establish a connection between artificial embryo cultivation and the successful implantation process.  Bioprinting in reproductive research: 3D bioprinting uses bioinks made from cells combined with hydrogels and ECM components to build tissue layers one by one. The reproductive technology application of this technology involves three uses i.e., foliculogenesis takes place on printed ovarian scaffolds, imitation of individual endometrial environments through printing for implantation studies and the technology allows for enduring hormonesensitive tissue development. Bioprinting technology helps create species-specific reproductive models that veterinary ART and conservation biology benefit from as reported (Heo et al., 2010; Laronda et al., 2017) [17,25] .  Organ on a Chips and Bioprinting present multiple advantages for ART procedures: These testing platforms are beneficial for evaluating embryo culture media while also assessing toxicant effects and individualized fertility treatments.

Advancements, challenges and future directions in microfluidics for assisted reproductive technologies

The evolution of microfluidics technology led to the development of comprehensive reproductive platforms which now perform automated gamete handling (Lai et al., 2015) [24] , The platform performs in vitro fertilization and tracks embryos (Heo et al., 2012) [17] . Previous studies used real-time metabolic monitoring techniques. Hormoneresponsive organ-on-a-chip models for ovarian and uterine tissues (Gargus et al., 2020; De Bem et al) [14]. [26] . Significant progress has been achieved but multiple key limitations remain unaddressed. Despite major advances in microfluidic-assisted reproductive technologies, several important challenges continue to limit their large-scale clinical and veterinary application. One of the primary concerns is biocompatibility, particularly in polydimethylsiloxane (PDMS)-based devices, as PDMS can absorb small hydrophobic molecules including steroid hormones and toxicants, thereby altering media composition and cellular responses (Toepke and Beebe, 2006; Berthier et al., 2012) [43, 3] . Another major limitation is workflow integration, because most IVF laboratories still rely on manual and highly variable protocols, making integration of automated microfluidic systems challenging within existing ART procedures (Swain et al., 2013) [42] . In addition, the lack of standardized fabrication methods, chip geometries, and validation criteria creates difficulties in reproducibility and inter-laboratory comparisons (Whitesides, 2006) [44] . Some advanced platforms also require specialized technical expertise and clean-room fabrication facilities, restricting their routine adoption in reproductive laboratories (Sackmann et al., 2014) [35] . Furthermore, species-specific optimization remains limited in veterinary ART, as many currently available devices are primarily designed for human reproductive systems and have not yet been fully validated for bovine or porcine applications (Ferraz et al., 2018) [12] . The integration of on-chip sensors to track embryo pH levels and oxygen consumption and glucose concentration in real-time enables non-invasive embryo viability predictions. AI and Machine Learning The combination of embryo development metrics with AI technology will enhance embryo selection performance especially when it uses time-lapse imaging (Simopoulou et al., 2018) [37] . Portable microfluidic devices that use sperm assessment on farms or in patients' homes will transform livestock reproductive technology and fertility testing. Patient-derived endometrial or ovarian cells in bioprinted chips could help scientists develop individualized fertility treatments and sustainable materials the development of bioinert and biodegradable chips will solve issues related to environmental and biological toxicity.

Conclusions

Microfluidic technology has emerged as a transformative tool in assisted reproductive technologies by enabling precise, automated, and physiologically relevant manipulation of gametes and embryos. Compared with conventional ART procedures, microfluidic systems provide superior control over fluid dynamics, nutrient exchange, temperature gradients, and cellular microenvironments while minimizing mechanical and oxidative stress. The integration of sperm sorting platforms, microfluidic IVF systems, embryo culture devices, and cryopreservation modules has significantly improved the efficiency and reproducibility of in vitro embryo production. Recent advances in three-dimensional culture systems, organ-on-achip platforms, and reproductive bioprinting have further expanded the scope of microfluidics by enabling realistic simulation of ovarian, endometrial, and embryo-maternal interactions. These biomimetic systems provide valuable opportunities for studying reproductive physiology, implantation biology, toxicology, and personalized fertility treatments in both human and veterinary medicine. Despite substantial progress, several challenges remain, including device standardization, large-scale manufacturing, biocompatibility of materials, integration with existing laboratory workflows, and species-specific optimization for veterinary applications. Future developments involving artificial intelligence, biosensors and portable lab-on-chip platforms are expected to accelerate the clinical and fieldlevel adoption of microfluidic ART systems. Overall, the convergence of microfluidics with reproductive biotechnology represents a promising next-generation approach for improving fertility management, embryo production, and reproductive research in animals and humans.

Acknowledgement

AcknowledgementsThe authors extend their gratitude to the ICAR-National Dairy Research Institute, Karnal, for providing the necessary support. Special thanks to colleagues and staff for their valuable insights and technical assistance.

References

1. Aydin S, Yaşlı M, Yildiz Ş, Urman B. Advancements in three-dimensional bioprinting for reproductive tissue engineering. Reprod Biomed Online. 2024;49(2):103118. https://doi.org/10.1016/j.rbmo.2024.04.002.
2. Bahat A, Eisenbach M. Sperm thermotaxis. Mol Cell Endocrinol. 2006;252(1-2):115-119. https://doi.org/10.1016/j.mce.2006.03.027.
3. Berthier E, Young EWK, Beebe DJ. Engineers are from PDMS-land, biologists are from polystyrenia. Lab Chip. 2012;12(7):1224-1237. https://doi.org/10.1039/C2LC20982A.
4. Bhat GR, Lone FA, Dalal J. Microfluidics-A novel technique for high-quality sperm selection for greater International Journal of Advanced Biochemistry Research https://www.biochemjournal.com ART outcomes. FASEB BioAdv. 2024;6:406-423. https://doi.org/10.1096/fba.2024-00041.
5. Bhatia SN, Ingber DE. Microfluidic organs-on-chips. Nat Biotechnol. 2014;32(8):760-772. https://doi.org/10.1038/nbt.2989.
6. Cho BS, Schuster TG, Zhu X, Chang D, Smith GD, Takayama S. Passively driven integrated microfluidic system for separation of motile sperm. Anal Chem. 2003;75(7):1671-1675. https://doi.org/10.1021/ac020579e.
7. Eisenbach M, Giojalas LC. Sperm guidance in mammals-an unpaved road to the egg. Nat Rev Mol Cell Biol. 2006;7(4):276-285. https://doi.org/10.1038/nrm1893.
8. Eisenbach M. Sperm navigation in humans: A concerted action of multiple guidance mechanisms. Commun Biol. 2025;8:923. https://doi.org/10.1038/s42003-025-08358-4.
9. El-Sherry TM, Abdelgawad M, Hussein HA, AbdElhafee H. Rheotaxis of sperm in fertile and infertile men. Syst Biol Reprod Med. 2023;69(1):1-11. https://doi.org/10.1080/19396368.2022.2141154.
10. Ferraz MAMM, Ferronato GA. Opportunities involving microfluidics and 3D culture systems for in vitro embryo production. Anim Reprod. 2023;20(2):e20230058. https://doi.org/10.1590/19843143-AR2023-0058.
11. Ferraz MAMM, Henning HHW, Stout TAE, Vos PLAM, Gadella BM. Designing three-dimensional in vitro oviduct culture systems to study mammalian fertilization and embryo production. Ann Biomed Eng. 2017;45(7):1731-1744. https://doi.org/10.1007/s10439016-1760-x.
12. Ferraz MAMM, Rho HS, Hemerich D, et al. An oviduct-on-a-chip provides an enhanced in vitro environment for zygote genome reprogramming. Nat Commun. 2018;9:4934. https://doi.org/10.1038/s41467018-07119-8.
13. Gadelha H, Cosson J, Smith DJ, Kirkman-Brown J. Human sperm uses asymmetric and anisotropic flagellar controls to regulate swimming symmetry and cell steering. eLife. 2019;8:e43714. https://doi.org/10.7554/eLife.43714.
14. Gargus ES, Tuan RS, Chen B. Engineering reproductive tissues and organoids using microfluidic systems. Reprod Biol Endocrinol. 2020;18:114. https://doi.org/10.1186/s12958-020-00674-4.
15. Gurkan UA, Wood DK, Carranza D, et al. Fulfilling the promise of lab-on-a-chip technologies. Lab Chip. 2024;24:1867-1874. https://doi.org/10.1039/D3LC00796K.
16. Han C, Zhang Q, Ma R, et al. Integration of single oocyte trapping, in vitro fertilization, and embryo culture in a microfluidic device. Lab Chip. 2010;10(21):2848-2854. https://doi.org/10.1039/C004954K.
17. Heo YS, Cabrera LM, Song JW, et al. Dynamic microfunnel culture enhances mouse embryo development and pregnancy rates. Hum Reprod. 2010;25(3):613-622. https://doi.org/10.1093/humrep/dep449.
18. Hoffman BD, Grashoff C, Schwartz MA. Dynamic molecular processes mediate cellular mechanotransduction. Nature. 2011;475(7356):316323. https://doi.org/10.1038/nature10316.
19. Iranzo AG, Vicente-Carrillo A, Ariza M, et al. Sperm selection using microfluidic techniques significantly improves sperm quality compared with conventional methods. Front Cell Dev Biol. 2025;13:1607927. https://doi.org/10.3389/fcell.2025.1607927.
20. Jahangiri AR, Ziarati N, Dadkhah E, et al. Microfluidics: The future of sperm selection in assisted reproduction. Andrology. 2024;12:1236-1252. https://doi.org/10.1111/andr.13578.
21. Jensen C, Teng Y. Is it time to start transitioning from 2D to 3D cell culture? Front Mol Biosci. 2020;7:33. https://doi.org/10.3389/fmolb.2020.00033.
22. Kantsler V, Dunkel J, Blayney M, Goldstein RE. Rheotaxis facilitates upstream navigation of mammalian sperm cells. eLife. 2014;3:e02403. https://doi.org/10.7554/eLife.02403.
23. Khodamoradi M, Rafizadeh Tafti S, Mousavi Shaegh SA, et al. Recent microfluidic innovations for sperm sorting. Chemosensors. 2021;9(6):126. https://doi.org/10.3390/chemosensors9060126.
24. Lai D, Takayama S, Smith GD. Recent microfluidic devices for studying gamete and embryo physiology. J Assist Reprod Genet. 2015;32(5):707-719. https://doi.org/10.1007/s10815-015-0452-9.
25. Laronda MM, Rutz AL, Xiao S, et al. A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice. Nat Commun. 2017;8:15261. https://doi.org/10.1038/ncomms15261.
26. Lee JB, Park JY, Kim SK, et al. Microfluidic organ-ona-chip platforms for reproductive biology and assisted reproductive technologies. Bioengineering. 2022;9(11):662. https://doi.org/10.3390/bioengineering9110662.
27. Li Z, Liu W, Qiu T, et al. The construction of an interfacial valve-based microfluidic chip for thermotaxis evaluation of human sperm. Biomicrofluidics. 2014;8(2):024102. https://doi.org/10.1063/1.4866851.
28. Ma J, Xie Q, Zhang Y, et al. Advances in microfluidic technology for sperm screening and in vitro fertilization. Anal Bioanal Chem. 2024;416:3717-3735. https://doi.org/10.1007/s00216-023-05120-9.
29. Miki K, Clapham DE. Rheotaxis guides mammalian sperm. Curr Biol. 2013;23(6):443-452. https://doi.org/10.1016/j.cub.2013.02.007.
30. Nasr-Esfahani MH, Deemeh MR, Tavalaee M, et al. Zeta sperm selection improves pregnancy rate and alters sex ratio in male factor infertility patients: a doubleblind randomized clinical trial. Int J Fertil Steril. 2016;10(2):253-260. https://doi.org/10.22074/ijfs.2016.4918.
31. Nosrati R, Vollmer M, Eamer L, et al. Rapid selection of sperm with high DNA integrity. Lab Chip. 2014;14(6):1142-1150. https://doi.org/10.1039/C3LC51254A.
32. Pensabene V, Agate F, Santos Miranda A, Picton HM. Microfluidics for in vitro fertilization: From science to clinical translation. Hum Reprod Update. 2026;32(2):206-230. https://doi.org/10.1093/humupd/dmaf028. International Journal of Advanced Biochemistry Research https://www.biochemjournal.com
33. Ricci G, Perticarari S, Boscolo R, et al. Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique. Fertil Steril. 2009;91(2):632-638. https://doi.org/10.1016/j.fertnstert.2007.
34. Richards HA, Eastman AJ, Miller DR, Cliffel DE. Reproductive organ-on-a-chip technologies and assessments of the fetal-maternal interface. Front Lab Chip Technol. 2024.
35. Sackmann EK, Fulton AL, Beebe DJ. The present and future role of microfluidics in biomedical research. Nature. 2014;507(7491):181-189. https://doi.org/10.1038/nature13118.
36. Samuel R, Feng H, Jafek A, et al. Microfluidic-based sperm sorting and analysis for treatment of male infertility. Methods Mol Biol. 2018;1748:197-217. https://doi.org/10.1007/978-1-4939-7694-0_16.
37. Simopoulou M, Sfakianoudis K, Maziotis E, et al. Are computational applications the "crystal ball" in the IVF laboratory? J Assist Reprod Genet. 2018;35(9):15451557. https://doi.org/10.1007/s10815-018-1278-6.
38. Sittadjody S, Saul JM, McQuilling JP, et al. In vivo transplantation of 3D encapsulated ovarian constructs in rats corrects abnormalities of ovarian failure. Nat Commun. 2017;8:1858. https://doi.org/10.1038/s41467017-01851-3.
39. Squires TM, Quake SR. Microfluidics: fluid physics at the nanoliter scale. Rev Mod Phys. 2005;77(3):9771026. https://doi.org/10.1103/RevModPhys.77.977.
40. Stern-Tal S, Ackerman S, Ben-David U, Shachar M. Biomimetic platforms for studying embryo implantation and endometrial receptivity. Biomaterials. 2020;260:120328. https://doi.org/10.1016/j.biomaterials.2020.120328.
41. Stone HA, Stroock AD, Ajdari A. Engineering flows in small devices: microfluidics toward a lab-on-a-chip. Annu Rev Fluid Mech. 2004;36:381-411. https://doi.org/10.1146/annurev.fluid.36.050802.12212
42. Swain JE, Lai D, Takayama S, Smith GD. Thinking big by thinking small: application of microfluidic technology to improve ART. Lab Invest. 2013;93(1):25-38. https://doi.org/10.1038/labinvest.2012.156.
43. Toepke MW, Beebe DJ. PDMS absorption of small molecules and consequences in microfluidic applications. Lab Chip. 2006;6(12):1484-1486. https://doi.org/10.1039/B612140C.
44. Whitesides GM. The origins and the future of microfluidics. Nature. 2006;442(7101):368-373. https://doi.org/10.1038/nature05058.
45. Wu T, Yan J, Nie K, et al. Microfluidic chips in female reproduction: a systematic review of status, advances, and challenges. Theranostics. 2024;14(11):4352-4374. https://doi.org/10.7150/thno.97301.
46. Xiao S, Coppeta JR, Rogers HB, et al. A microfluidic culture model of the human reproductive tract and 28day menstrual cycle. Nat Commun. 2017;8:14584. https://doi.org/10.1038/ncomms14584.
47. Yanez LZ, Camarillo DB. Microfluidic devices for oocyte and embryo manipulation. Adv Drug Deliv Rev. 2017;114:116-131. https://doi.org/10.1016/j.addr.2017.03.009.
48. Yanez LZ, Han J, Behr BB, et al. Human oocyte developmental potential is predicted by mechanical properties within hours after fertilization. Nat Commun. 2016;7:10809. https://doi.org/10.1038/ncomms10809.
49. Zaferani M, Cheong SH, Abbaspourrad A. Rheotaxisbased separation of sperm with progressive motility using a microfluidic corral system. Proc Natl Acad Sci U S A. 2019;116(11):4897-4902. https://doi.org/10.1073/pnas.1817359116.
50. Zhang YS, Aleman J, Shin SR, et al. Multisensorintegrated organs-on-chips platform for automated and continual in situ monitoring of organoid behaviors. Proc Natl Acad Sci U S A. 2017;114(12):E2293-E2302. https://doi.org/10.1073/pnas.1612906114.
51. Zhao XM, Ren JJ, Du WH, et al. Effect of threedimensional culture in alginate hydrogel on developmental competence of bovine embryos. Zygote. 2015;23(5):706-715.