Single oocyte manipulation in microfluidic channels via precisely controlled flow is critical in microfluidics-based in vitro fertilization. Such systems can potentially minimize the number of transfer steps among containers for rinsing as often performed during conventional in vitro fertilization and can standardize protocols by minimizing manual handling steps. To study shape deformation of oocytes under shear flow and its subsequent impact on their spindle structure is essential for designing microfluidics for in vitro fertilization. Here, we developed a simple yet powerful approach to (1) trap a single oocyte and induce its deformation through a constricted microfluidic channel, (2) quantify oocyte deformation in real time using a conventional microscope and (3) retrieve the oocyte from the microfluidic device to evaluate changes in their spindle structures. We found that oocytes can be significantly deformed under high flow rates, e.g., 10 μL/min in a constricted channel with a width and height of 50 and 150 μm, respectively. Oocyte spindles can be severely damaged, as shown here by immunocytochemistry staining of the microtubules and chromosomes. The present approach can be useful to investigate underlying mechanisms of oocyte deformation exposed to well-controlled shear stresses in microfluidic channels, which enables a broad range of applications for reproductive medicine.
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Abkarian M, Faivre M, Viallat A (2007) Swinging of red blood cells under shear flow. Phys Rev Lett 98:188302
Asghar W, Velasco V, Kingsley JL, Shoukat MS, Shafiee H, Anchan RM, Demirci U (2014) Selection of functional human sperm with higher DNA integrity and fewer reactive oxygen species. Adv Healthc Mater. doi:10.1002/adhm.201400058
Assal RE, Guven S, Gurkan UA, Gozen I, Shafiee H, Dalbeyler S, Abdalla N, Thomas G, Fuld W, Illigens BMW, Estanislau J, Khoory J, Kaufman R, Zylberberg C, Lindeman N, Wen Q, Ghiran I, Demirci U (2014) Bio‐Inspired Cryo‐Ink Preserves Red Blood Cell Phenotype and Function During Nanoliter Vitrification. Adv Mater. doi:10.1002/adma.201400941
Clark SG, Haubert K, Beebe DJ, Ferguson CE, Wheeler MB (2005) Reduction of polyspermic penetration using biomimetic microfluidic technology during in vitro fertilization. Lab Chip 5:1229–1232
Deschamps J, Kantsler V, Segre E, Steinberg V (2009) Dynamics of a vesicle in general flow. Proc Natl Acad Sci USA 106:11444–11447
Dumoulin JCM, Coonen E, Bras M, Bergers-Janssen JM, Ignoul-Vanvuchelen RCM, van Wissen LCP, Geraedts JPM, Evers JLH (2001) Embryo development and chromosomal anomalies after ICSI: effect of the injection procedure. Hum Reprod 16:306–312
Ebner T, Moser M, Sommergruber M, Puchner M, Wiesinger R, Tews G (2003) Developmental competence of oocytes showing increased cytoplasmic viscosity. Hum Reprod 18:1294–1298
El-Ali J, Sorger PK, Jensen KF (2006) Cells on chips. Nature 442:403–411
Gurkan UA, Tasoglu S, Akkaynak D, Avci O, Unluisler S, Canikyan S, MacCallum N, Demirci U (2012) Smart interface materials integrated with microfluidics for on-demand local capture and release of cells. Adv Healthc Mater 1:661–668
Han C, Zhang Q, Ma R, Xie L, Qiu T, Wang L, Mitchelson K, Wang J, Huang G, Qiao J, Cheng J (2010) Integration of single oocyte trapping, in vitrofertilization and embryo culture in a microwell-structured microfluidic device. Lab Chip 10:2848–2854
Heo YS, Lee H-J, Hassell BA, Irimia D, Toth TL, Elmoazzen H, Toner M (2011) Controlled loading of cryoprotectants (CPAs) to oocyte with linear and complex CPA profiles on a microfluidic platform. Lab Chip 11:3530–3537
Hu L-L, Shen X-H, Zheng Z, Wang Z-D, Liu Z-H, Jin L-H, Lei L (2012) Cytochalasin B treatment of mouse oocytes during intracytoplasmic sperm injection (ICSI) increases embryo survival without impairment of development. Zygote 20:361–369
Jimenez AM, Roche M, Pinot M, Panizza P, Courbin L, Gueroui Z (2011) Towards high throughput production of artificial egg oocytes using microfluidics. Lab Chip 11:429–434
Kim T, Wang CW, Thomas FIM, Sastry AM (2006) Fluid-structure interaction analysis of flow-induced deformation in a two- phase, Neo-Hookean marine egg. J Eng Mater-T ASME 128:519–526
Lathi RB, Milki AA (2004) Rate of aneuploidy in miscarriages following in vitro fertilization and intracytoplasmic sperm injection. Fertil Steril 81:1270–1272
Liu X, Fernandes R, Jurisicova A, Casper RF, Sun Y (2010) In situ mechanical characterization of mouse oocytes using a cell holding device. Lab Chip 10:2154–2161
Luo ZY, Wang SQ, He L, Lu TJ, Xu F, Bai BF (2013) Front tracking simulation of cell detachment dynamic mechanism in microfluidics. Chem Eng Sci 97:394–405
Navarro P, Liu L, Trimarchi JR, Ferriani RA, Keefe DL (2005) Noninvasive imaging of spindle dynamics during mammalian oocyte activation. Fertil Steril 83:1197–1205
Papi M, Brunelli R, Sylla L, Parasassi T, Monaci M, Maulucci G, Missori M, Arcovito G, Ursini F, De Spirito M (2010) Mechanical properties of zona pellucida hardening. Eur Biophys J Biophy 39:987–992
Raju GAR, Prakash GJ, Krishna KM, Madan K (2007) Meiotic spindle and zona pellucida characteristics as predictors of embryonic development: a preliminary study using PolScope imaging. Reprod Biomed Online 14:166–174
Rizvi I, Gurkan UA, Tasoglu S, Alagic N, Celli JP, Mensah LB, Mai Z, Demirci U, Hasan T (2013) Flow induces epithelial-mesenchymal transition, cellular heterogeneity and biomarker modulation in 3D ovarian cancer nodules. Proc Natl Acad Sci USA 110:E1974–E1983
Sadani Z, Wacogne B, Pieralli C, Roux C, Gharbi T (2005) Microsystems and microfluidic device for single oocyte transportation and trapping: toward the automation of in vitro fertilising. Sensor Actuat A-Phys 121:364–372
Suh RS, Zhu XY, Phadke N, Ohl DA, Takayama S, Smith GD (2006) IVF within microfluidic channels requires lower total numbers and lower concentrations of sperm. Hum Reprod 21:477–483
Sun QY, Schatten H (2006) Regulation of dynamic events by microfilaments during oocyte maturation and fertilization. Reproduction 131:193–205
Sun Y, Wan KT, Roberts KP, Bischof JC, Nelson BJ (2003) Mechanical property characterization of mouse zona pellucida. IEEE Trans Nanobiosci 2:279–286
Tasoglu S, Gurkan UA, Wang S, Demirci U (2013a) Manipulating biological agents and cells in micro-scale volumes for applications in medicine. Chem Soc Rev 42:5788–5808
Tasoglu S, Safaee H, Zhang X, Kingsley JL, Catalano PN, Gurkan UA, Nureddin A, Kayaalp E, Anchan RM, Maas RL, Tuezel E, Demirci U (2013b) Exhaustion of racing sperm in nature-mimicking microfluidic channels during sorting. Small 9:3374–3384
Tasoglu S, Diller E, Guven S, Sitti M, Demirci U (2014) Untethered micro-robotic coding of three-dimensional material composition. Nat Commun 5:3124
Thomas FIM, Bolton TF (1999) Shear stress experienced by echinoderm eggs in the oviduct during spawning: potential role in the evolution of egg properties. J Exp Biol 202:3111–3119
Vanapalli SA, Duits MHG, Mugele F (2009) Microfluidics as a functional tool for cell mechanics. Biomicrofluidics 3:012006
Wang WH, Meng L, Hackett RJ, Odenbourg R, Keefe DL (2001) Limited recovery of meiotic spindles in living human oocytes after cooling-rewarming observed using polarized light microscopy. Hum Reprod 16:2374–2378
Zeggari R, Wacogne B, Pieralli C, Roux C, Gharbi T (2007) Microfluidic applications for andrology. Sensor Actuat B-Chem 125:664–671
Zenzes MT, Bielecki R, Casper RF, Leibo SP (2001) Effects of chilling to 0 degrees C on the morphology of meiotic spindles in human metaphase II oocytes. Fertil Steril 75:769–777
Zhang X, Khimji I, Shao L, Safaee H, Desai K, Keles HO, Gurkan UA, Kayaalp E, Nurreddin A, Anchan RM, Maas RL, Demirci U (2012) Nanoliter droplet vitrification for oocyte cryopreservation. Nanomedicine 7:553–564
Utkan Demirci acknowledges that this work is supported by the National Institutes of Health award (R01 EB015776). We would like to acknowledge Dr. Aida Nureddin for her significant contributions to this work. ZhengYuan Luo acknowledges financial supports from China Scholarship Council. This work was also partially supported by Swedish Research Council Vetenskapsrådet.
Conflict of interest
Dr. Utkan Demirci is a founder of and has an equity interest in: (1) DxNow Inc., a company that is developing microfluidic and imaging technologies for point-of-care diagnostic solutions, and (2) Koek Biotech, a company that is developing microfluidic IVF technologies for clinical solutions. Dr. Utkan Demirci’s interests were viewed and managed by the conflict of interest policies.
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Luo, Z., Güven, S., Gozen, I. et al. Deformation of a single mouse oocyte in a constricted microfluidic channel. Microfluid Nanofluid 19, 883–890 (2015). https://doi.org/10.1007/s10404-015-1614-0
- Oocyte deformation
- Spindle damage
- Single cell trapping