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

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.	周新麗, 衣星越, 周楠峰, 等. 用于低溫保護劑混合的微流控芯片設計及優化. 生物醫學工程學雜志, 2016, 35(3): 461-465.
18.	戴建軍. 冷凍保存對豬卵母細胞凋亡模式與基因表達的影響. 南京: 南京農業大學, 2015.
19.	張德福, 朱良成, 劉東, 等. 豬卵母細胞冷凍保存研究. 中國農業科學, 2006, 39(6): 1233-1240.
20.	楊喜, 吳靜, 劉丑生, 等. 細胞松弛素 B、溫度、離心處理對豬卵母細胞 OPS 玻璃化冷凍的影響. 內蒙古大學學報: 自然科學版, 2013(5): 525-529.
21.	田見暉, 劉國世, 曾申明, 等. 電脈沖激活對體外成熟豬卵母細胞卵裂率和囊胚率的影響. 農業生物技術學報, 2004, 12(4): 401-407.
22.	趙金鳳. 豬卵母細胞孤雌激活方法的研究. 哈爾濱: 東北農業大學, 2010.
23.	郝子悅. 豬卵母細胞體外成熟及孤雌激活的研究. 揚州: 揚州大學, 2008.
24.	羅婷, 蒙麗娜, 劉曉華, 等. 哺乳動物附植前胚胎細胞凋亡的研究進展. 中國畜牧獸醫, 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. 海藻糖的性質及應用. 中國生化藥物雜志, 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. 周新麗, 衣星越, 周楠峰, 等. 用于低溫保護劑混合的微流控芯片設計及優化. 生物醫學工程學雜志, 2016, 35(3): 461-465.
18. 戴建軍. 冷凍保存對豬卵母細胞凋亡模式與基因表達的影響. 南京: 南京農業大學, 2015.
19. 張德福, 朱良成, 劉東, 等. 豬卵母細胞冷凍保存研究. 中國農業科學, 2006, 39(6): 1233-1240.
20. 楊喜, 吳靜, 劉丑生, 等. 細胞松弛素 B、溫度、離心處理對豬卵母細胞 OPS 玻璃化冷凍的影響. 內蒙古大學學報: 自然科學版, 2013(5): 525-529.
21. 田見暉, 劉國世, 曾申明, 等. 電脈沖激活對體外成熟豬卵母細胞卵裂率和囊胚率的影響. 農業生物技術學報, 2004, 12(4): 401-407.
22. 趙金鳳. 豬卵母細胞孤雌激活方法的研究. 哈爾濱: 東北農業大學, 2010.
23. 郝子悅. 豬卵母細胞體外成熟及孤雌激活的研究. 揚州: 揚州大學, 2008.
24. 羅婷, 蒙麗娜, 劉曉華, 等. 哺乳動物附植前胚胎細胞凋亡的研究進展. 中國畜牧獸醫, 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. 海藻糖的性質及應用. 中國生化藥物雜志, 1995(5): 231-233.

Journal of Biomedical Engineering

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

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