Morphofunctional Characteristics of Freshly Isolated and Cryopreserved Human Ovarian Granulosa and Cumulus Cells

Authors

  • Marina P. Petrushko Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov
  • Vladimir I. Pinyaev Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov
  • Elena B. Revenko Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov
  • Natalia A. Volkova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov
  • Natalia N. Chub Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov

DOI:

https://doi.org/10.15407/cryo24.01.057

Keywords:

granulosa, cumulus, culture, cryopreservation

Abstract

Utilization of granulosa and cumulus cell (GCC) complex could be prospective for co-culture of gametes and embryos as a part of assisted reproductive technologies (ART), that makes important their cryopreservation. The study was performed in GCCs of women aged 25–39, who underwent the treatment of infertility by IVF. Cryopreservation of GCC suspension was carried out using 1.5 M 1,2-propane diol solution. The samples were cooled with rate of 0.3 deg/min from 25 down to –6°C, thereafter the ice crystal initiation was done, from –6 down to –35°C the cooling rate was 1 deg/min, after that the samples were plunged into liquid nitrogen and stored then at –196°C. Following thawing the GCCs retained their main features: ability for adhesion and proliferation, and hormone production. During culture of GCCs its development was delayed for 1–2 days if compared with freshly isolated cells. In 7 days after explanting the cultures of frozen-thawed GCCs appeared nearly the same as primary cultures of corresponding terms of in vitro culture. The post thaw level of hormone production by GCCs was changed: decreased by 89.9% in case of estradiol, and increased by 81.4% for progesterone if compared with the indices of freshly isolated suspensions. The obtained results testify to a possibility to isolate the GCCs and then to store in low-temperature banks for prospective use in ART.


Probl Cryobiol Cryomed 2014; 24(1):57–66.

Author Biographies

Marina P. Petrushko, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov

Department of Cryobiology of Reproduction System

Vladimir I. Pinyaev, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov

Department of Cryobiology of Reproduction System

Elena B. Revenko, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov

Department of Cryobiology of Reproduction System

Natalia A. Volkova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov

Department of Cryobiochemistry

Natalia N. Chub, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkov

Department of Cryobiology of Reproduction System

References

Alisch A., Ruping K., Koster F. et al. Cumulus cell apoptosis as a predictor for oocyte quality in artificial reproduction technique. Zentralbl Gynakol 2003; 125(11): 452–457. PubMed

Castro S.V., de Carvalho A.A., da Silva C.M., et al. Freezing solution containing dimethylsulfoxide and fetal calf serum maintains survival and ultrastructure of goat preantral follicles after cryopreservation and in vitro culture of ovarian tissue. Cell Tissue Res 2011; 346(2): 283–292. CrossRef PubMed

Chub N.N., Lobyntseva G.S., Demina L.G. Effect of cryoprotectants on morphological and functional integrity of human ovarian tissue. In: Cryopreservation of cells and tissues. Kharkov; 1989: p. 118. PubMed

Isachenko V., Isachenko E., Mallmann P., Rahimi G. Increasing follicular and stromal cell proliferation in cryopreserved human ovarian tissue after long-term precooling prior to freezing: in vitro versus chorioallantoic membrane (CAM) xenotransplantation. Cell Transplant 2013; 22(11): 2053–2061. CrossRef PubMed

Freeman M., Whitworth C., Hill G. Granulosa cell co-culture enhances human embryo development and pregnancy rate following in vitro fertilization. Hum Reprod 1995; 10(2): 408–414. PubMed

Fuller B., Green C., Grischenko V.I. Cryopreservation for cell banking: current concepts at the turn of 21st century. Problems of Cryobiology 2003; (2): 62–83.

Grischenko V.I., Paraschuk Yu.S., Dakhno F.V., Yurchenko G.G. Cryobiology and infertility problems. Kiev: Naukova Dumka; 1990.

Gunasena K., Villines P., Critser J. Live births after autologous transplantation of cryopreserved mouse ovaries. Hum Reprod 1997; 12(1): 101–106. CrossRef PubMed

Gook D.A., Osborn S.M., Bourne H., Johnston W.I. Fertilization of human oocytes following cryopreservation; normal karyotypes and absence of stray chromosomes. Hum Reprod 1994; 9(4): 684–691. PubMed

Gook D.A., Osborn S.M., Johnston W.I. Cryopreservation of mouse and human oocytes using 1,2-propanediol and the configuration of the meiotic spindle. Hum Reprod 1993; 8(7): 1101–1109. PubMed

Gook D.A., Osborn S.M., Johnston W.I. Parthenogenetic activation of human oocytes following cryopreservation using 1,2-propanediol. Hum Reprod 1995; 10(3): 654–658. PubMed

Gook D.A., Schiewe M.C., Osborn S.M. et al. Intracytoplasmic sperm injection and embryo development of human oocytes cryopreserved using 1,2-propanediol. Hum Reprod 1995; 10(10): 2637–2641. PubMed

Host E., Mikkelsen A.L., Lindenberg S., Smidt-Jensen S. Apoptosis in human cumulus cells in relation to maturation stage and cleavage of the corresponding oocyte. Acta Obstet Gynecol Scand 2000; 79(11): 936–940. PubMed

Johnson J., Higdon H., Boone W. Effect of human granulosa cell co-culture using standard culture media on the maturation and fertilization potential of immature human oocytes. Fertil Steril 2008; 90(5): 1674–1679. CrossRef PubMed

Lindley E., Jacobson J., Corselli J. Cryopreservation of human cumulus cells for co-cultures and assessment of DNA damage after thawing using the comet assay II. J Assist Reprod Genet 2001; 18(10): 534–538. CrossRef PubMed

Liu Y., Holyoak G., Wang S., Bunch T. The importance of cumulus cells on the in vitro production of bovine oocytes. Theriogenology 1995; 43(1): 267. CrossRef

Lebedeva I. Yu., Kibardina T.V., Kuzmina T.I. Participation of granulosa cells in mediating effect of prolactin and somatotropin on bovine oocyte-cumulus complexes in vitro. Tsytologiya 2005; 47(10): 882–888.

McNatty K.P., Baird D.T., Bolton A. et al. Concentration of estrogens and androgens in human ovarian venous plasma and follicular fluid throughout the menstrual cycle. J Endocrinol 1976; 71(2): 77–85. CrossRef PubMed

Muiheron G., Bossert N., Lapp J. et al. Human granulose-luteal and cumulus cells ezpress transforming growth factors-beta type 1 and type 2 mRNA. J Clin Endocrinol Metab 1992; 74(2): 32–40.

Parikh F., Nadkarni S., Naik N. еt al. Cumulus coculture and cumulus-aided embryo transfer increases pregnancy rates in patients undergoing in vitro fertilization. Fertil Steril 2006; 86(4): 839–847. CrossRef PubMed

Petrenko A.Y. Study of mitochondria membrane reparation after freeze-thawing. Kriobiologiya 1987; (2): 24–29.

Ruppert-Lingham C.J., Paynter S.J., Godfrey J. et al. Developmental potential of murine germinal vesicle stage cumulus-oocyte complexes following exposure to dimethylsulphoxide or cryo-preservation: loss of membrane integrity of cumulus cells after thawing. Hum Reprod 2003; 18(2): 392–398. CrossRef PubMed

Zhang A., Xu B., Sun Y. et al. The effect of human cumulus cells on the maturation and developmental potential of immature oocytes in ICSI cycles. J Assist Reprod Genet 2012; 29(4): 313–319. CrossRef PubMed

Downloads

Published

2014-03-25

How to Cite

Petrushko, M. P., Pinyaev, V. I., Revenko, E. B., Volkova, N. A., & Chub, N. N. (2014). Morphofunctional Characteristics of Freshly Isolated and Cryopreserved Human Ovarian Granulosa and Cumulus Cells. Problems of Cryobiology and Cryomedicine, 24(1), 57–66. https://doi.org/10.15407/cryo24.01.057

Issue

Section

Theoretical and Experimental Cryobiology