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Sadat Tahajjodi, S., Farashahi Yazd, E., Agha-Rahimi, A., Aflatoonian, R., Ali Khalili, M., Mohammadi, M., & Aflatoonian, B. (2020). Biological and physiological characteristics of human cumulus cell in adherent culture condition. International Journal of Reproductive BioMedicine (IJRM), 18(1), 1-10. https://doi.org/10.18502/ijrm.v18i1.6189
https://doi.org/10.18502/ijrm.v18i1.6189
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Somayyeh Sadat Tahajjodi
Somayyeh Sadat Tahajjodi
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
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Ehsan Farashahi Yazd
Ehsan Farashahi Yazd
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
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Azam Agha-Rahimi
Azam Agha-Rahimi
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
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Reza Aflatoonian
Reza Aflatoonian
Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Centre, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
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Mohammad Ali Khalili
Mohammad Ali Khalili
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
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Mahnaz Mohammadi
Mahnaz Mohammadi
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
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Behrouz Aflatoonian
Behrouz Aflatoonian
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Advanced Medical Sciences and Technologies, School of Paramedicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Published Date
- Jan 26, 2020
Biological and physiological characteristics of human cumulus cell in adherent culture condition
Abstract
Background: Cumulus cells, as oocyte nurse cells, provide a suitable microenvironment with growth factors and cellular interactions required for oocyte maturation. Thus, these cells may serve as a natural niche for in vitro studies of female germ cell development. Cumulus cells may help attain a better understanding of the causes of infertility in women and eventually improve the outcomes of cases that respond poorly to standard infertility treatment.
Objective: The aim of this study was to isolate, culture, and investigate the biological characteristics of human cumulus cells.
Materials and Methods: In this experimental study, cumulus cells were isolated, cultured, and characterized using reverse transcription-polymerase chain reaction analyses of specific genes including FOXL2, CYP19A1, FSHR, AMHR, and LHR. The presence of vimentin, a structural protein, was examined via immunofluorescent staining. Moreover, levels of anti-mullerian hormone (AMH) and progesterone secretion by cumulus cells were measured with ELISA after 2, 4, 12, 24, and 48 hr of culture.
Results: In adherent culture, human cumulus cells expressed specific genes and markers as well as secreted AMH and progesterone into the medium.
Conclusion: Cumulus cells secrete AMH and progesterone in an adherent culture and might be applicable for in vitro maturation (IVM) and in vitro gametogenesis (IVG) studies.
Key words: Cumulus cells, Conditioned medium, In vitro maturation, In vitro gametogenesis, Niche.
References
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[2] Assou S, Pourret E, Péquignot M, Rigau V, Kalatzis V, Ait-Ahmed O, et al. Cultured cells from the human oocyte cumulus niche are efficient feeders to propagate pluripotent stem cells. Stem Cells Dev 2015; 24: 2317–2327.
[3] Huang Z, Wells D. The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol Hum Reprod 2010; 16: 715–725.
[4] Chand AL, Legge M. Amino acid transport system L activity in developing mouse ovarian follicles. Hum Reprod 2011; 26: 3102–3108.
[5] Siristatidis CS, Vrachnis N, Creatsa M, Maheshwari A, Bhattacharya S. In vitro maturation in subfertile women with polycystic ovarian syndrome undergoing assisted reproduction. Cochrane Database Syst Rev 2013; 10: CD006606.
[6] Lee HJ, Quaas AM, Wright DL, Toth TL, Teixeira JM. In vitro maturation (IVM) of murine and human germinal vesicle (GV)-stage oocytes by coculture with immortalized human fallopian tube epithelial cells. Fertil Steril 2011; 95: 1344–1348.
[7] Ben-Ami I, Komsky A, Bern O, Kasterstein E, Komarovsky D, Ron-El R. In vitro maturation of human germinal vesicle-stage oocytes: role of epidermal growth factor-like growth factors in the culture medium. Hum Reprod 2011; 26: 76–81.
[8] Feng G, Shi D, Yang S, Wang X. Co-culture embedded in cumulus clumps promotes maturation of denuded oocytes and reconstructs gap junctions between oocytes and cumulus cells. Zygote 2013; 21: 231–237.
[9] Zhou CJ, Wu SN, Shen JP, Wang DH, Kong XW, Lu A, et al. The beneficial effects of cumulus cells and oocyte-cumulus cell gap junctions depends on oocyte maturation and fertilization methods in mice. Peer J 2016; 4: e1761.
[10] Fatehi AN, Roelen BA, Colenbrander B, Schoevers EJ, Gadella BM, Bevers MM, et al. Presence of cumulus cells during in vitro fertilization protects the bovine oocyte against oxidative stress and improves first cleavage but does not affect further development. Zygote 2005; 13: 177–185.
[11] Wongsrikeao P, Kaneshige Y, Ooki R, Taniguchi M, Agung B, Nii M, et al. Effect of the removal of cumulus cells on the nuclear maturation, fertilization and development of porcine oocytes. Reprod Domest Anim 2005; 40: 166–170.
[12] Ju S, Rui R. Effects of cumulus cells on in vitro maturation of oocytes and development of cloned embryos in the pig. Reprod Domest Anim 2012; 47: 521–529.
[13] Zhang A, Xu B, Sun Y, Lu X, Niu Z, Chen Q, 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: 313–319.
[14] Madkour A, Bouamoud N, Kaarouch I, Louanjli N, Saadani B, Assou S, et al. Follicular fluid and supernatant from cultured cumulus-granulosa cells improve in vitro maturation in patients with polycystic ovarian syndrome. Fertil Steril 2018; 110: 710–719.
[15] Vithoulkas A, Levanduski M, Goudas VT, Illmensee K. Co-culture of human embryos with autologous cumulus cell clusters and its beneficial impact of secreted growth factors on preimplantation development as compared to standard embryo culture in assisted reproductive technologies (ART). Middle East Fertility Society Journal 2017; 22: 317– 322.
[16] Cihangir N, Görkemli H, Özdemir S, Aktan M, Duman S. Influence of cumulus cell coculture and cumulusaided embryo transfer on embryonic development and pregnancy rates. J Turk Ger Gynecol Assoc 2010; 11: 121–126.
[17] Bhadarka HK, Patel NH, Patel NH, Patel M, Patel KB, Sodagar NR, et al. Impact of embryo co-culture with cumulus cells on pregnancy & implantation rate in patients undergoing in vitro fertilization using donor oocyte. Indian J Med Res 2017; 146: 341–345.
[18] Abdel Ghani MA, Abe Y, Asano T, Hamano S, Suzuki H. Effect of bovine cumulus-oocyte complexes conditioned medium on in vitro maturation of canine oocytes. Reprod Med Biol 2011; 10: 43–49.
[19] Chen HF, Jan PS, Kuo HC, Wu FC, Lan CW, Huang MC, et al. Granulosa cells and retinoic acid cotreatment enrich potential germ cells from manually selected Oct4-EGFP expressing human embryonic stem cells. Reprod Biomed Online 2014; 29: 319–332.
[20] Qing T, Shi Y, Qin H, Ye X, Wei W, Liu H, et al. Induction of oocyte like cells from mouse embryonic stem cells by co-culture with ovarian granulosa cells. Differentiation 2007; 75: 902–911.
[21] Sadeghian-Nodoushan F, Aflatoonian R, Borzouie Z, Akyash F, Fesahat F, Soleimani M, et al. Pluripotency and differentiation of cells from human testicular sperm extraction: An investigation of cell stemness. Mol Reprod Dev 2016; 83: 312–323.
[22] Leung DTH, Fuller PJ, Chu S. Impact of FOXL2 mutations on signaling in ovarian granulosa cell tumors. Int J Biochem Cell Biol 2016; 72: 51–54.
[23] Yang YJ, Wang Y, Li Z, Zhou L, Gui JF. Sequential, divergent and cooperative requirements of Foxl2a and Foxl2b in ovary development and maintenance of zebrafish. Genetics 2017; 205: 1551–1572.
[24] Hosseini E, Mehraein F, Shahhoseini M, Karimian L, Nikmard F, Ashrafi M, et al. Epigenetic alterations of CYP19A1 gene in Cumulus cells and its relevance to infertility in endometriosis. J Assist Reprod Genet 2016; 33: 1105–1113.
[25] Sacchi S, Marinaro F, Xella S, Marsella T, Tagliasacchi D, La Marca A. The anti-Müllerian hormone (AMH) induces forkhead box L2 (FOXL2) expression in primary culture of human granulosa cells in vitro. J Assist Reprod Genet 2017; 34: 1131–1136.
[26] Jeppesen JV, Kristensen SG, Nielsen ME, Humaidan P, Dal Canto M, Fadini R, et al. LH-receptor gene expression in human granulosa and cumulus cells from antral and preovulatory follicles. J Clin Endocrinol Metab 2012; 97: E1524–E1531.
[27] Karakaya C, Guzeloglu-Kayisli O, Hobbs RJ, Gerasimova T, Uyar A, Erdem M, et al. Folliclestimulating hormone receptor (FSHR) alternative skipping of exon 2 or 3 affects ovarian response to FSH. Mol Hum Reprod 2014; 20: 630–643.
[28] Dewailly D, Robin G, Peigne M, Decanter C, Pigny P, Catteau-Jonard S. Interactions between androgens, FSH, anti-Müllerian hormone and estradiol during folliculogenesis in the human normal and polycystic ovary. Hum Reprod Update 2016; 22: 709–724.
[29] Kusaka CS, Utsunomiya T, Kumasako Y, Otsu E, Mori T, Shimada M. The relationship between the level of progesterone secreted from cumulus cells and oocyte developmental competence in in vitro matured human cumulus oocyte complexes. J Mamm Ova Res 2012; 29: 41–47.
[30] Nagyova E, Scsukova S, Kalous J, Mlynarcikova A. Effects of RU486 and indomethacin on meiotic maturation, formation of extracellular matrix, and progesterone production by porcine oocytecumulus complexes. Domest Anim Endocrinol 2014; 48: 7–14.
[31] Kempisty B, Ziółkowska A, Ciesiółka S, Piotrowska H, Antosik P, Bukowska D, et al. Expression and cellular distribution of estrogen and progesterone receptors and the real-time proliferation of porcine cumulus cells. Zygote 2015; 23: 836–845.
[32] Kedem A, Yung Y, Yerushalmi GM, Haas J, Maman E, Hanochi M, et al. Anti mullerian hormone (AMH) level and expression in mural and cumulus cells in relation to age. J Ovarian Res 2014; 7: 113.
[33] Convissar S, Armouti M, Fierro MA, Winston NJ, Scoccia H, Zamah AM, et al. Regulation of amhby oocyte specific growth factors in human primary cumulus cells. Reproduction 2017; 154: 745–753.
[34] Grøndahl ML, Nielsen ME, Dal Canto M, Fadini R, Rasmussen IA, Westergaard LG, et al. Anti- Müllerian hormone remains highly expressed in human cumulus cells during the final stages of folliculogenesis. Reprod Biomed Online 2011; 22: 389–398.
[35] Zhang Y, Shao L, Xu Y, Cui Y, Liu J, Chian RC. Effect of anti-Mullerian hormone in culture medium on quality of mouse oocytes matured in vitro. PLoS One 2014; 9: e99393.
[2] Assou S, Pourret E, Péquignot M, Rigau V, Kalatzis V, Ait-Ahmed O, et al. Cultured cells from the human oocyte cumulus niche are efficient feeders to propagate pluripotent stem cells. Stem Cells Dev 2015; 24: 2317–2327.
[3] Huang Z, Wells D. The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol Hum Reprod 2010; 16: 715–725.
[4] Chand AL, Legge M. Amino acid transport system L activity in developing mouse ovarian follicles. Hum Reprod 2011; 26: 3102–3108.
[5] Siristatidis CS, Vrachnis N, Creatsa M, Maheshwari A, Bhattacharya S. In vitro maturation in subfertile women with polycystic ovarian syndrome undergoing assisted reproduction. Cochrane Database Syst Rev 2013; 10: CD006606.
[6] Lee HJ, Quaas AM, Wright DL, Toth TL, Teixeira JM. In vitro maturation (IVM) of murine and human germinal vesicle (GV)-stage oocytes by coculture with immortalized human fallopian tube epithelial cells. Fertil Steril 2011; 95: 1344–1348.
[7] Ben-Ami I, Komsky A, Bern O, Kasterstein E, Komarovsky D, Ron-El R. In vitro maturation of human germinal vesicle-stage oocytes: role of epidermal growth factor-like growth factors in the culture medium. Hum Reprod 2011; 26: 76–81.
[8] Feng G, Shi D, Yang S, Wang X. Co-culture embedded in cumulus clumps promotes maturation of denuded oocytes and reconstructs gap junctions between oocytes and cumulus cells. Zygote 2013; 21: 231–237.
[9] Zhou CJ, Wu SN, Shen JP, Wang DH, Kong XW, Lu A, et al. The beneficial effects of cumulus cells and oocyte-cumulus cell gap junctions depends on oocyte maturation and fertilization methods in mice. Peer J 2016; 4: e1761.
[10] Fatehi AN, Roelen BA, Colenbrander B, Schoevers EJ, Gadella BM, Bevers MM, et al. Presence of cumulus cells during in vitro fertilization protects the bovine oocyte against oxidative stress and improves first cleavage but does not affect further development. Zygote 2005; 13: 177–185.
[11] Wongsrikeao P, Kaneshige Y, Ooki R, Taniguchi M, Agung B, Nii M, et al. Effect of the removal of cumulus cells on the nuclear maturation, fertilization and development of porcine oocytes. Reprod Domest Anim 2005; 40: 166–170.
[12] Ju S, Rui R. Effects of cumulus cells on in vitro maturation of oocytes and development of cloned embryos in the pig. Reprod Domest Anim 2012; 47: 521–529.
[13] Zhang A, Xu B, Sun Y, Lu X, Niu Z, Chen Q, 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: 313–319.
[14] Madkour A, Bouamoud N, Kaarouch I, Louanjli N, Saadani B, Assou S, et al. Follicular fluid and supernatant from cultured cumulus-granulosa cells improve in vitro maturation in patients with polycystic ovarian syndrome. Fertil Steril 2018; 110: 710–719.
[15] Vithoulkas A, Levanduski M, Goudas VT, Illmensee K. Co-culture of human embryos with autologous cumulus cell clusters and its beneficial impact of secreted growth factors on preimplantation development as compared to standard embryo culture in assisted reproductive technologies (ART). Middle East Fertility Society Journal 2017; 22: 317– 322.
[16] Cihangir N, Görkemli H, Özdemir S, Aktan M, Duman S. Influence of cumulus cell coculture and cumulusaided embryo transfer on embryonic development and pregnancy rates. J Turk Ger Gynecol Assoc 2010; 11: 121–126.
[17] Bhadarka HK, Patel NH, Patel NH, Patel M, Patel KB, Sodagar NR, et al. Impact of embryo co-culture with cumulus cells on pregnancy & implantation rate in patients undergoing in vitro fertilization using donor oocyte. Indian J Med Res 2017; 146: 341–345.
[18] Abdel Ghani MA, Abe Y, Asano T, Hamano S, Suzuki H. Effect of bovine cumulus-oocyte complexes conditioned medium on in vitro maturation of canine oocytes. Reprod Med Biol 2011; 10: 43–49.
[19] Chen HF, Jan PS, Kuo HC, Wu FC, Lan CW, Huang MC, et al. Granulosa cells and retinoic acid cotreatment enrich potential germ cells from manually selected Oct4-EGFP expressing human embryonic stem cells. Reprod Biomed Online 2014; 29: 319–332.
[20] Qing T, Shi Y, Qin H, Ye X, Wei W, Liu H, et al. Induction of oocyte like cells from mouse embryonic stem cells by co-culture with ovarian granulosa cells. Differentiation 2007; 75: 902–911.
[21] Sadeghian-Nodoushan F, Aflatoonian R, Borzouie Z, Akyash F, Fesahat F, Soleimani M, et al. Pluripotency and differentiation of cells from human testicular sperm extraction: An investigation of cell stemness. Mol Reprod Dev 2016; 83: 312–323.
[22] Leung DTH, Fuller PJ, Chu S. Impact of FOXL2 mutations on signaling in ovarian granulosa cell tumors. Int J Biochem Cell Biol 2016; 72: 51–54.
[23] Yang YJ, Wang Y, Li Z, Zhou L, Gui JF. Sequential, divergent and cooperative requirements of Foxl2a and Foxl2b in ovary development and maintenance of zebrafish. Genetics 2017; 205: 1551–1572.
[24] Hosseini E, Mehraein F, Shahhoseini M, Karimian L, Nikmard F, Ashrafi M, et al. Epigenetic alterations of CYP19A1 gene in Cumulus cells and its relevance to infertility in endometriosis. J Assist Reprod Genet 2016; 33: 1105–1113.
[25] Sacchi S, Marinaro F, Xella S, Marsella T, Tagliasacchi D, La Marca A. The anti-Müllerian hormone (AMH) induces forkhead box L2 (FOXL2) expression in primary culture of human granulosa cells in vitro. J Assist Reprod Genet 2017; 34: 1131–1136.
[26] Jeppesen JV, Kristensen SG, Nielsen ME, Humaidan P, Dal Canto M, Fadini R, et al. LH-receptor gene expression in human granulosa and cumulus cells from antral and preovulatory follicles. J Clin Endocrinol Metab 2012; 97: E1524–E1531.
[27] Karakaya C, Guzeloglu-Kayisli O, Hobbs RJ, Gerasimova T, Uyar A, Erdem M, et al. Folliclestimulating hormone receptor (FSHR) alternative skipping of exon 2 or 3 affects ovarian response to FSH. Mol Hum Reprod 2014; 20: 630–643.
[28] Dewailly D, Robin G, Peigne M, Decanter C, Pigny P, Catteau-Jonard S. Interactions between androgens, FSH, anti-Müllerian hormone and estradiol during folliculogenesis in the human normal and polycystic ovary. Hum Reprod Update 2016; 22: 709–724.
[29] Kusaka CS, Utsunomiya T, Kumasako Y, Otsu E, Mori T, Shimada M. The relationship between the level of progesterone secreted from cumulus cells and oocyte developmental competence in in vitro matured human cumulus oocyte complexes. J Mamm Ova Res 2012; 29: 41–47.
[30] Nagyova E, Scsukova S, Kalous J, Mlynarcikova A. Effects of RU486 and indomethacin on meiotic maturation, formation of extracellular matrix, and progesterone production by porcine oocytecumulus complexes. Domest Anim Endocrinol 2014; 48: 7–14.
[31] Kempisty B, Ziółkowska A, Ciesiółka S, Piotrowska H, Antosik P, Bukowska D, et al. Expression and cellular distribution of estrogen and progesterone receptors and the real-time proliferation of porcine cumulus cells. Zygote 2015; 23: 836–845.
[32] Kedem A, Yung Y, Yerushalmi GM, Haas J, Maman E, Hanochi M, et al. Anti mullerian hormone (AMH) level and expression in mural and cumulus cells in relation to age. J Ovarian Res 2014; 7: 113.
[33] Convissar S, Armouti M, Fierro MA, Winston NJ, Scoccia H, Zamah AM, et al. Regulation of amhby oocyte specific growth factors in human primary cumulus cells. Reproduction 2017; 154: 745–753.
[34] Grøndahl ML, Nielsen ME, Dal Canto M, Fadini R, Rasmussen IA, Westergaard LG, et al. Anti- Müllerian hormone remains highly expressed in human cumulus cells during the final stages of folliculogenesis. Reprod Biomed Online 2011; 22: 389–398.
[35] Zhang Y, Shao L, Xu Y, Cui Y, Liu J, Chian RC. Effect of anti-Mullerian hormone in culture medium on quality of mouse oocytes matured in vitro. PLoS One 2014; 9: e99393.