Effect of cryoprotectant removal by microfluidic chip on developmental capacity of oocytes_Journal of Biomedical Engineering

Authors：

YI Xingyue ¹ ,  ZHOU Xinli ¹ , YANG Yun ¹ , DAI Jianjun ² , ZHANG Defu ²

1. Institute of Biothermal Technology, University of Shanghai for Science and Technology, Shanghai 200093, P.R.China;
2. Animal and Veterinary Research Institute, SAAS, Shanghai 201106, P.R.China;

Corresponding author：

ZHOU Xinli, Email: zjulily@163.com

Keywords：

cryoprotectants; microfluidic chip; oocytes; dilution protocol

DOI：

10.7507/1001-5515.201608014

Video：

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In order to reduce osmotic damage and chemical toxicity of cryoprotectants (CPA) to oocytes during unloading process, the microfluidic chip was used to remove CPA from porcine MⅡ oocytes in this study. Firstly, the effects of unloading time, composition and concentration of diluting solutions of microfluidic method on survival rate and developmental capacity of oocytes were studied, then microfluidic method was compared with traditional one-step and two-step CPA unloading protocols. The results showed that when the total time is 8 minutes, the survival rate and morula rate of oocytes treated with microfluidic method could achieve 95.99% ± 4.64% and 74.17% ± 1.18%, respectively, which were not significantly different from fresh control group (98.53% ± 2.94%; 78.22% ± 1.34%). In addition, 1 mol/L sucrose diluting solutions were more beneficial than other solutions, and it was also showed that microfluidic method achieved better survival, cleavage rate of oocytes than traditional methods. Microfluidic CPA removal protocol can reduce the damage to oocytes during unloading process, and may further improve the cryopreservation effect of oocytes.

Citation： YI Xingyue, ZHOU Xinli, YANG Yun, DAI Jianjun, ZHANG Defu. Effect of cryoprotectant removal by microfluidic chip on developmental capacity of oocytes. Journal of Biomedical Engineering, 2018, 35(1): 123-130. doi: 10.7507/1001-5515.201608014 Copy

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2.	al-Hasani S, Kirsch J, Diedrich K, et al. Successful embryo transfer of cryopreserved and in-vitro fertilized rabbit oocytes. Hum Reprod, 1989, 4(1): 77-79.
3.	Fuku E, Kojimat T, Shioya Y, et al. In vitro fertilization and development of frozen-thawed bovine oocytes. Cryobiology, 1992, 29(4): 485-492.
4.	Maclellan L J, Carnevale E M, Coutinho da Silva M A, et al. Pregnancies from vitrified equine oocytes collected from super-stimulated and non-stimulated mares. Theriogenology, 2002, 58(5): 911-919.
5.	Chen H H, Shen Hong, Heimfeld S, et al. A microfluidic study of mouse dendritic cell membrane transport properties of water and cryoprotectants. Int J Heat Mass Transf, 2008, 51(23/24): 5687-5694.
6.	Kobel S, Valero A, Latt J, et al. Optimization of microfluidic single cell trapping for long-term on-chip culture. Lab Chip, 2010, 10(7): 857-863.
7.	Lecault V, White A K, Singhal A, et al. Microfluidic single cell analysis: from promise to practice. Curr Opin Chem Biol, 2012, 16(3/4): 381-390.
8.	Song Y S, Moon S, Hulli L, et al. Microfluidics for cryopreservation. Lab Chip, 2009, 9(13): 1874-1881.
9.	Hubel A. Advancing the preservation of cellular therapy products. Transfusion, 2011, 51(Suppl 4): 82S-86S.
10.	Fleming K K, Longmire E K, Hubel A. Numerical characterization of diffusion-based extraction in cell-laden flow through a microfluidic channel. J Biomech Eng, 2007, 129(5): 703-711.
11.	Mata C, Longmire E, Mckenna D, et al. Cell motion and recovery in a two-stream microfluidic device. Microfluid Nanofluidics, 2010, 8(4): 457-465.
12.	Mata C, Longmire E K, Mckenna D H, et al. Experimental study of diffusion-based extraction from a cell suspension. Microfluid Nanofluidics, 2008, 5(4): 529-540.
13.	Hanna J, Hubel A, Lemke E. Diffusion-based extraction of DMSO from a cell suspension in a three stream, vertical microchannel. Biotechnol Bioeng, 2012, 109(9): 2316-2324.
14.	Heo Y S, Lee H J, Hassell B A, et al. Controlled loading of cryoprotectants (CPAs) to oocyte with linear and complex CPA profiles on a microfluidic platform. Lab Chip, 2011, 11(20): 3530-3537.
15.	Paynter S J, O'neil L, Fuller B J, et al. Membrane permeability of human oocytes in the presence of the cryoprotectant propane-1,2-diol. Fertil Steril, 2001, 75(3): 532-538.
16.	Paynter S J, Borini A, Bianchi V, et al. Volume changes of mature human oocytes on exposure to cryoprotectant solutions used in slow cooling procedures. Hum Reprod, 2005, 20(5): 1194-1199.
17.	周新丽, 衣星越, 周楠峰, 等. 用于低温保护剂混合的微流控芯片设计及优化. 生物医学工程学杂志, 2016, 35(3): 461-465.
18.	戴建军. 冷冻保存对猪卵母细胞凋亡模式与基因表达的影响. 南京: 南京农业大学, 2015.
19.	张德福, 朱良成, 刘东, 等. 猪卵母细胞冷冻保存研究. 中国农业科学, 2006, 39(6): 1233-1240.
20.	杨喜, 吴静, 刘丑生, 等. 细胞松弛素 B、温度、离心处理对猪卵母细胞 OPS 玻璃化冷冻的影响. 内蒙古大学学报: 自然科学版, 2013(5): 525-529.
21.	田见晖, 刘国世, 曾申明, 等. 电脉冲激活对体外成熟猪卵母细胞卵裂率和囊胚率的影响. 农业生物技术学报, 2004, 12(4): 401-407.
22.	赵金凤. 猪卵母细胞孤雌激活方法的研究. 哈尔滨: 东北农业大学, 2010.
23.	郝子悦. 猪卵母细胞体外成熟及孤雌激活的研究. 扬州: 扬州大学, 2008.
24.	罗婷, 蒙丽娜, 刘晓华, 等. 哺乳动物附植前胚胎细胞凋亡的研究进展. 中国畜牧兽医, 2012, 39(11): 125-128.
25.	Martino A, Songsasen N, Leibo S P. Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. Biol Reprod, 1996, 54(5): 1059-1069.
26.	Crowe J H, Carpenter J F, Crowe L M. The role of vitrification in anhydrobiosis. Annu Rev Physiol, 1998, 60(1): 73-103.
27.	李群, 袁勤生. 海藻糖的性质及应用. 中国生化药物杂志, 1995(5): 231-233.

1. Chen C. Pregnancy after human oocyte cryopreservation. Lancet, 1986, 1(8486): 884-886.
2. al-Hasani S, Kirsch J, Diedrich K, et al. Successful embryo transfer of cryopreserved and in-vitro fertilized rabbit oocytes. Hum Reprod, 1989, 4(1): 77-79.
3. Fuku E, Kojimat T, Shioya Y, et al. In vitro fertilization and development of frozen-thawed bovine oocytes. Cryobiology, 1992, 29(4): 485-492.
4. Maclellan L J, Carnevale E M, Coutinho da Silva M A, et al. Pregnancies from vitrified equine oocytes collected from super-stimulated and non-stimulated mares. Theriogenology, 2002, 58(5): 911-919.
5. Chen H H, Shen Hong, Heimfeld S, et al. A microfluidic study of mouse dendritic cell membrane transport properties of water and cryoprotectants. Int J Heat Mass Transf, 2008, 51(23/24): 5687-5694.
6. Kobel S, Valero A, Latt J, et al. Optimization of microfluidic single cell trapping for long-term on-chip culture. Lab Chip, 2010, 10(7): 857-863.
7. Lecault V, White A K, Singhal A, et al. Microfluidic single cell analysis: from promise to practice. Curr Opin Chem Biol, 2012, 16(3/4): 381-390.
8. Song Y S, Moon S, Hulli L, et al. Microfluidics for cryopreservation. Lab Chip, 2009, 9(13): 1874-1881.
9. Hubel A. Advancing the preservation of cellular therapy products. Transfusion, 2011, 51(Suppl 4): 82S-86S.
10. Fleming K K, Longmire E K, Hubel A. Numerical characterization of diffusion-based extraction in cell-laden flow through a microfluidic channel. J Biomech Eng, 2007, 129(5): 703-711.
11. Mata C, Longmire E, Mckenna D, et al. Cell motion and recovery in a two-stream microfluidic device. Microfluid Nanofluidics, 2010, 8(4): 457-465.
12. Mata C, Longmire E K, Mckenna D H, et al. Experimental study of diffusion-based extraction from a cell suspension. Microfluid Nanofluidics, 2008, 5(4): 529-540.
13. Hanna J, Hubel A, Lemke E. Diffusion-based extraction of DMSO from a cell suspension in a three stream, vertical microchannel. Biotechnol Bioeng, 2012, 109(9): 2316-2324.
14. Heo Y S, Lee H J, Hassell B A, et al. Controlled loading of cryoprotectants (CPAs) to oocyte with linear and complex CPA profiles on a microfluidic platform. Lab Chip, 2011, 11(20): 3530-3537.
15. Paynter S J, O'neil L, Fuller B J, et al. Membrane permeability of human oocytes in the presence of the cryoprotectant propane-1,2-diol. Fertil Steril, 2001, 75(3): 532-538.
16. Paynter S J, Borini A, Bianchi V, et al. Volume changes of mature human oocytes on exposure to cryoprotectant solutions used in slow cooling procedures. Hum Reprod, 2005, 20(5): 1194-1199.
17. 周新丽, 衣星越, 周楠峰, 等. 用于低温保护剂混合的微流控芯片设计及优化. 生物医学工程学杂志, 2016, 35(3): 461-465.
18. 戴建军. 冷冻保存对猪卵母细胞凋亡模式与基因表达的影响. 南京: 南京农业大学, 2015.
19. 张德福, 朱良成, 刘东, 等. 猪卵母细胞冷冻保存研究. 中国农业科学, 2006, 39(6): 1233-1240.
20. 杨喜, 吴静, 刘丑生, 等. 细胞松弛素 B、温度、离心处理对猪卵母细胞 OPS 玻璃化冷冻的影响. 内蒙古大学学报: 自然科学版, 2013(5): 525-529.
21. 田见晖, 刘国世, 曾申明, 等. 电脉冲激活对体外成熟猪卵母细胞卵裂率和囊胚率的影响. 农业生物技术学报, 2004, 12(4): 401-407.
22. 赵金凤. 猪卵母细胞孤雌激活方法的研究. 哈尔滨: 东北农业大学, 2010.
23. 郝子悦. 猪卵母细胞体外成熟及孤雌激活的研究. 扬州: 扬州大学, 2008.
24. 罗婷, 蒙丽娜, 刘晓华, 等. 哺乳动物附植前胚胎细胞凋亡的研究进展. 中国畜牧兽医, 2012, 39(11): 125-128.
25. Martino A, Songsasen N, Leibo S P. Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. Biol Reprod, 1996, 54(5): 1059-1069.
26. Crowe J H, Carpenter J F, Crowe L M. The role of vitrification in anhydrobiosis. Annu Rev Physiol, 1998, 60(1): 73-103.
27. 李群, 袁勤生. 海藻糖的性质及应用. 中国生化药物杂志, 1995(5): 231-233.

Journal of Biomedical Engineering

Effect of cryoprotectant removal by microfluidic chip on developmental capacity of oocytes

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