Influence of programmable cryopreservation and in vitro culture on follicular viability and estradiol secretion of human ovarian tissue

doi:10.3969/j.issn.1006-7795.2013.04.006

Abstract

Abstract:

Objective To investigate the influence of programmable cryopreservation and in vitro culture on follicular viability and estradiol secretion of human ovarian tissue. Methods Pieces of human ovarian cortex were prepared, the specimens were round in shape and were about 1 mm×1 mm×1 mm, and were divided into fresh group, frozen group, fresh culturing group and freezing-thawing culturing group. Pieces of human ovarian cortex were frozen with the method of programmable cryopreservation and cultured in vitro before freezing and after thawing. The level of estradiol in medium was measured with electro-chemiluminescence immunoassay. Healthy follicles stained with Clacein AM were conducted under fluorescence microscopy after digestion for all ovarian tissues. Results The number of follicles and their proportions in freezing group had no significant difference from that in fresh group(P>0.05). Estradiol was secreted continuously throughout in-vitro culture in both of fresh tissues and freezing-thawing tissues. The level of estradiol in freezing-thawing culturing group was lower than that in fresh culturing group in the first 2 d(P<0.05). The difference of the estradiol level was not significant(P>0.05) after 2 d. The number of primordial and primary follicles and their proportions had no significant difference after in vitro culture(P>0.05); the number of secondary follicles and its proportion were higher in tissues cultured in vitro for 14 days(P<0.05). Conclusion The method of programmable cryopreservation can maintain the viability of preantral follicles and E₂ secretion of ovarian tissues cultured in vitro. Secondary follicles have greater potential of development than that of primordial and primary follicles of ovarian tissues cultured in vitro.

Key words: ovarian tissue, programmable cryopreservation, in vitro culture

CLC Number:

R339.2

TIAN Xuanxuan, RUAN Xiangyan, Montag Markus, Liebenthron Jana, Alfred O. Mueck. Influence of programmable cryopreservation and in vitro culture on follicular viability and estradiol secretion of human ovarian tissue[J]. Journal of Capital Medical University, 2013, 34(4): 506-511.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2013.04.006

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2013/V34/I4/506

References

[1] Smitz J, Dolmans M M, Donnez, J, et al. Current achievements and future culture, in vitro follicle development and transplantation: implications for fertility preservation[J]. Human Reproduction Update, 2010,16(4):395-414.

[2] Bedaiwy M A, Falcone T. Harvesting and autotransplantation of vascularized ovarian grafts: approaches and techniques[J]. Reprod Biomed Online, 2007,14(3):360-371.

[3] Gougeon A. Dynamics of follicular growth in the human: a model from preliminary results[J]. Hum Reprod, 1986,1(2):81-87.

[4] Gosden R G, Baird D T, Wade J C, et al. Restoration of fertility to oophorectomized sheep by ovarian autografts stored at-96 degrees C[J]. Hum Reprod, 1994,9(4):597-603.

[5] Michael V W, Jacques D, Outi H, et al. Cryopreservation and autotransplantation of human ovarian tissue prior to cytotoxic therapy-A technique in its infancy but already successful in fertility preservation[J]. European Journal of Cancer, 2009,45(9):1547-1553.

[6] Silber S J. Ovary cryopreservation and transplantation for fertility preservation[J]. Molecular Human Reproduction, 2012,18(2):59-67.

[7] 曹金燕,史小林,诸定寿.大鼠胚胎卵巢异体异位移植的探讨[J].首都医科大学学报,2000,21(1):9-11.

[8] 李云秀,马艳萍,李永刚,等.不同冷冻方案对人类卵巢组织形态学的影响[J].中国优生与遗传杂志,2011,19(6):101-103.

[9] Dahl S L, Chen Z, Solan A K, et al. Feasibility of vitrification as a storage method for tissue-engineered blood vessels[J]. Tissue Eng, 2006,12(2):291-300.

[10] Fabbri R, Pasquinelli G, Bracone G. Cryopreservation of human ovarian tissue[J]. Cell Tissue Bank, 2006,7(2):123-133.

[11] Choi J, Lee B, Lee E, et al. Cryopreservation of ovarian tissues temporarily suppresses the proliferation of granulosa cells in mouse preantral follicles[J]. Cryobiology, 2008,56(1):36-42.

[12] Li Y B, Zhou C Q, Yang G F, et al. Modified vitrification method for cryopreservation of human ovarian tissues[J]. Chin Med, 2007,120(2):110-114.

[13] Bishonga C, Takahashi Y, Katagiri S, et al. In vitro growth of mouse ovarian preantral follicles and the capacity of their oocytes to develop to the blastocyst stage[J]. J Vet Med Sci, 2001,63(6):619-624.

[14] Yu Y, Li W, Han Z, et al. The effect of follicle-stimulating hotmone on follicular development granulosa cell apoptosis and stemidogenesis and its mediation by insulin-like growth factor-I in the goat ovary[J]. Theriogenology, 2003,60(9):1691-1704.

[15] Yang P, Roy S K. A novel mechanism of FSH regulation of DNA synthesis in the granulesa cell of hamster preantral follicles: involvement of a protein kinase C-mediated MAP kinase 3/1 selfactivation loop[J]. Biol Reprod, 2006,75(1):149-157.

[16] Brito A B, Santos R R, R. van den Hurk, et al. Short-term culture of ovarian cortical strips from capuchin monkeys: a morphological, viability, and molecular study of preantral follicular development in vitro[J]. Reproductive Sciences Published Online, 11 January 2013.

[17] Seema P, Deepa B, Dhananjay D M, et al. Stimulation of ovarian stem cells by follicle timulating hormone and basic fibroblast growth factor during cortical tissue culture[J]. J Ovarian Res, 2013,6:20.