Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.21451/1984-3143-AR2018-0083
Animal Reproduction (AR)
Original Article

Evaluation of stemness marker expression in bovine ovarian granulosa cells

Son Nghia Hoang, Chi Nguyen Quynh Ho, Thao Thi Phuong Nguyen, Chung Chinh Doan, Diem Hong Tran, Long Thanh Le

Downloads: 1
Views: 1394

Abstract

The objective of this study was to assess the stemness marker expressions (Oct4, Nanog, and Sox2) of granulosa cells (GCs) collected from bovine ovarian follicles and in vitro expansion. The single bovine ovarian follicles were isolated and categorized into 4 groups according to their diameter including group A (<2 mm), group B (2-3 mm), group C (3-4 mm), and group D (>4 mm). Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and immunostaining were applied to evaluate the stemness marker expression of bovine GCs from ovarian follicles. We also estimated the stemness marker transcript expressions of GCs during in vitro expression by qRT-PCR. qRT-PCR analysis demonstrated that fresh GCs from bovine ovarian follicles expressed the stemness markers (Oct4, Nanog, Sox2). These markers were down-regulated during antral stage follicular development. We also estimated stemness marker transcript expressions of GCs which were isolated and in vitro expanded from ovarian follicles of group A. The qRT-PCR results showed that Oct4 and Sox2 transcript expressions were reduced during in vitro expansion while Nanog transcript was not expressed.

Keywords

granulosa cells, ovarian follicles, transcript expression, stemness markers.

References

Bagheripour N, Zavareh S, Ghorbanian MT, Paylakhi SH, Mohebbi SR. 2017. Changes in the expression of OCT4 in mouse ovary during estrous cycle. Vet Res Forum, 8(1):43-48.

Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, Gifford DK, Melton DA, Jaenisch R, and Young RA. 2005. Core Transcriptional Regulatory Circuitry in Human Embryonic Stem Cells. Cell, 122(6):947-956.

Bui HT, Van Thuan N, Kwon DN, Choi YJ, Kang MH, Han JW, Kim T, Kim JH. 2014. Identification and characterization of putative stem cells in the adult pig ovary. Development, 141:2235-2244.

Chronowska E. 2012. Stem cell characteristics of ovarian granulose cells. Ann Anim Sci, 12(2):151-157.

Copper GM. 1997. The cell, a molecular approach. Oxford University Press, London. pp.604-608.

Dzafic E, Stimpfel M, Virant-Klun I. 2013. Plasticity of granulosa cells: on the crossroad of stemness and transdifferentiation potential. J Assist Reprod Genet, 30:1255-1261.

Gong SP, Lee ST, Lee EJ, Kim DY, Lee G, Chi SG, Ryu BK, Lee CH, Yum KE, Lee HJ, Han JY, Tilly JL, Lim JM. 2010. Embryonic stem cell-like cells established by culture of adult ovarian cells in mice. Fertil Steril, 93:2594-2601.

Gougeon A. 1996. Regulation of ovarian follicular development in primates: facts and hypotheses. Endocr Rev, 17(2):121-155.

Kossowska-Tomaszczuk K, De Geyter C, De Geyter M, Martin I, Holzgreve W, Scherberich A, Zhang H. 2009. The multipotency of luteinizing granulosa cells collected from mature ovarian follicles. Stem Cells, 27:210-219.

Lavranos TC, Mathis JM, Latham SE, Kalionis B,

Shay JW, and Rodgers RJ. 1999. Evidence for Ovarian Granulosa Stem Cells: Telomerase Activity and Localization of the Telomerase Ribonucleic Acid Component in Bovine Ovarian Follicles. Biol Reprod, 61:358-366.

Li H, Chian RC. 2012. Follicular Development and Oocyte Growth. Development of In Vitro Maturation for Human Oocytes. Springer, 37-57.

Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using Real-Time Quantitative PCR and the 2-∆∆Ct Method Methods, 25:402-408.

Mattioli M, Gloria A, Turriani M, Berardinelli P, Russo V, Nardinocchi D, Curini V, Baratta M, Martignani E, Barboni B. 2012. Osteo-regenerative potential of ovarian granulosa cells: An in vitro and in vivo study. Theriogenology, 77:1425-1437.

Oki Y, Ono H, Motohashi T, Sugiura N, Nobusue H, Kano K. 2012. Dedifferentiated follicular granulosa cells derived from pig ovary can transdifferentiate into osteoblasts. Biochem J, 447:239-248.

Parte S, Bhartiya D, Telang J, Daithankar V, Salvi V, Zaveri K, Hinduja I. 2011. Detection, characterization, and spontaneous differentiation in vitro of very small embryonic-like putative stem cells in adult mammalian ovary.  Stem Cells Dev, 20(8):1451-1464.

Potten CS, Loeffler M. 1990. Stem cells: attributes, cycles, spirals, pitfalls and uncertainties. Lessons for and from the crypt. Development, 110:1001-1020.

Rodgers RJ, Lavranos TC, van Wezel IL, Irving-Rodgers HF. 1999. Development of the ovarian follicular epithelium. Mol Cell Endocrinol, 151(1-2):171-179.

Rodgers RJ, Irving-Rodgers HF, van Wezel IL, Krupa M, Lavranos TC. 2001. Dynamics of the membrana granulosa during expansion of the ovarian follicular antrum. Mol Cell Endocrinol, 171(1-2):41-48.

Sumer H, Liu J, Malaver-Ortega LF, Lim ML, Khodadadi K, Verma PJ. 2011. NANOG is a key factor for induction of pluripotency in bovine adult fibroblasts. J Anim Sci, 89:2708-2716.

Townson DH, Combelles CMH. 2012. Ovarian Follicular Atresia, Basic Gynecology. Some Related Issues. InTech, 44-76.

Varras M, Griva T, Kalles V, Akrivis C, Paparisteidis N. 2012. Markers of stem cells in human ovarian granulosa cells: is there a clinical significance in ART?. J Ovarian Res, 5:36.

5cebdcb80e88259e0ca63c0f animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections