Utilizing calcium alginate for the assessment of bone morphogenetic protein 15 induction effect on the differentiation of mesenchymal stem cell derived from human follicular fluid to oocyte-like structure

Moghadam, Ali Reza; Taheri Moghadam, Mahin; Saki, Ghasem; Nikbakht, Roshan

Utilizing calcium alginate for the assessment of bone morphogenetic protein 15 induction effect on the differentiation of mesenchymal stem cell derived from human follicular fluid to oocyte-like structure

Document Type : Original Article

Authors

¹ Cellular and Molecular Research Center; Department of Anatomical Science, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

² Department of Anatomical Science, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

³ Fertility, Infertility and Perinatology Center, Imam Khomeini Hospital, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

Abstract

Background: Follicular fluid (FF)-derived mesenchymal stem cells (MSCs) are possible new source of cells in the study of oogenesis and regenerative medicine. Several biomaterials have been used as scaffolds to mimic ovarian tissue stroma. Using good matrix is essential for increasing the cell survival rate, proliferation, and differentiation. However, no study has been performed to investigate the effects of BMP15 and calcium alginate hydrogel on the differentiation potential of FF-derived MSCs to oocyte-like structures (OLSs). Materials and Methods: In this work, FF MSCs, which were collected from women in routine in vitro fertilization procedure, were capsulated with 0.5% calcium alginate, and then the encapsulated cells were cultured in medium containing BMP15 for 2 weeks. Trypan blue staining was carried out to determine cell viability. Real-time polymerase chain reaction (PCR) and immunofluorescence (ICC) staining method were performed to characterize the expression of OCT4, Nanog, ZP2, and ZP3 genes and protein. The encapsulation process did not change the morphology and viability of the encapsulated cells. Results: Reverse-transcription-PCR and ICC showed that MSCs expressed germ line stem cell markers such as OCT4 and Nanog. After 4 days of culture, OLSs formed and expressed zona pellucida markers. OLSs at least reached 180–230 μm in diameter in the control and BMP15-treated groups. Finally, a reduction in the expression pattern of pluripotency and ZP markers was detected in the encapsulated cells cultured in the BMP15-supplemented medium. Conclusion: The three-dimensional alginate culture system seems to be a promising method of getting in vitro differentiation and development of ovarian cells, which could mimic the native ovarian condition.

Keywords

References

1.	Mascarenhas MN, Flaxman SR, Boerma T, Vanderpoel S, Stevens GA. National, regional, and global trends in infertility prevalence since 1990: A systematic analysis of 277 health surveys. PLoS Med 2012;9:e1001356.
2.	Martin JJ, Woods DC, Tilly JL. Implications and current limitations of oogenesis from female germline or oogonial stem cells in adult mammalian ovaries. Cells. 2019 Feb; 8 (2):93.
3.	Porras-Gómez TJ, Moreno-Mendoza NJ. Neo-oogenesis in mammals. Zygote 2017;25:404-22.
4.	Hoang SN, Ho CN, Nguyen TT, Doan CC, Tran DH, Le LT. Evaluation of stemness marker expression in bovine ovarian granulosa cells. Animal Reproduction. 2019 Jun; 16 (2):277-81.
5.	Zheng T, Hong W, Ye H, Hu C, Z. Mechanism for the decision of ovarian surface epithelial stem cells to undergo neo-oogenesis or ovarian tumorigenesis. Cell Physiol Biochem 2018;50:214-32.
6.	Kossowska-Tomaszczuk K, De Geyter C, De Geyter M, Martin I, Holzgreve W, Scherberich A, et al. The multipotency of luteinizing granulosa cells collected from mature ovarian follicles. Stem Cells 2009;27:210-9.
7.	Woods DC, Tilly JL. Isolation, characterization and propagation of mitotically active germ cells from adult mouse and human ovaries. Nat Protoc 2013;8:966-88.
8.	Costa JJN, Souza GB, Soares MAA, Ribeiro RP, van den Hurk R, Silva JRV. In vitro differentiation of primordial germ cells and oocyte-like cells from stem cells. Histol Histopathol 2018;33:121-32.
9.	Ding X, Liu G, Xu B, Wu C, Hui N, Ni X, et al. Human GV oocytes generated by mitotically active germ cells obtained from follicular aspirates. Sci Rep 2016;6:28218.
10.	Belli M, Shimasaki S. Molecular aspects and clinical relevance of GDF9 and BMP15 in ovarian function. Vitam Horm 2018;107:317-48.
11.	Lochab AK, Extavour CG. Bone morphogenetic protein (BMP) signaling in animal reproductive system development and function. Dev Biol 2017;427:258-69.
12.	Bayne RA, Donnachie DJ, Kinnell HL, Childs AJ, Anderson RA. BMP signalling in human fetal ovary somatic cells is modulated in a gene-specific fashion by GREM1 and GREM2. Mol Hum Reprod 2016;22:622-33.
13.	Prapa E, Vasilaki A, Dafopoulos K, Katsiani E, Georgoulias P, Messini CI, et al. Effect of anti-Müllerian hormone (AMH) and bone morphogenetic protein 15 (BMP-15) on steroidogenesis in primary-cultured human luteinizing granulosa cells through Smad5 signalling. J Assist Reprod Genet 2015;32:1079-88.
14.	Hussein TS, Froiland DA, Amato F, Thompson JG, Gilchrist RB. Oocytes prevent cumulus cell apoptosis by maintaining a morphogenic paracrine gradient of bone morphogenetic proteins. J Cell Sci 2005;118:5257-68.
15.	Persani L, Rossetti R, Di Pasquale E, Cacciatore C, Fabre SJ. The fundamental role of bone morphogenetic protein 15 in ovarian function and its involvement in female fertility disorders. Hum Reprod Update 2014;20:869-83.
16.	Ramirez G, Palomino J, Aspee K, De los Reyes M. GDF-9 and BMP-15 mRNA Levels in Canine Cumulus Cells Related to Cumulus Expansion and the Maturation Process. Animals. 2020 Mar; 10 (3):462.
17.	Luciano AM, Franciosi F, Modina SC, Lodde V. Gap junction-mediated communications regulate chromatin remodeling during bovine oocyte growth and differentiation through cAMP-dependent mechanism (s). Biol Reprod 2011;85:1252-9.
18.	Belli M, Vigone G, Merico V, Redi CA, Zuccotti M, Garagna S. Towards a 3D culture of mouse ovarian follicles. Int J Dev Biol 2012;56:931-7.
19.	Correia HH, Lima LF, Sousa FG, Ferreira AC, Cadenas J, Paes VM, et al. Activation of goat primordial follicles in vitro: Influence of alginate and ovarian tissue. Reprod Domest Anim 2020;55:105-9.
20.	Brito IR, Lima IM, Xu M, Shea LD, Woodruff TK, Figueiredo JR. Three-dimensional systems for in vitro follicular culture: Overview of alginate-based matrices. Reprod Fertil Dev 2014;26:915-30.
21.	Lee KY, Mooney DJ. Alginate: Properties and biomedical applications. Prog Polym Sci 2012;37:106-26.
22.	Sadr SZ, Fatehi R, Maroufizadeh S, Amorim CA, Ebrahimi B. Utilizing fibrin-alginate and Matrigel-alginate for mouse follicle development in three-dimensional culture systems. Biopreserv Biobank 2018;16:120-7.
23.	Sittadjody S, Saul JM, Joo S, Yoo JJ, Atala A, Opara EC. Engineered multilayer ovarian tissue that secretes sex steroids and peptide hormones in response to gonadotropins. Biomaterials 2013;34:2412-20.
24.	Riva F, Omes C, Bassani R, Nappi RE, Mazzini G, Icaro Cornaglia A, et al. In vitro culture system for mesenchymal progenitor cells derived from waste human ovarian follicular fluid. Reprod Biomed Online 2014;29:457-69.
25.	Lai D, Xu M, Zhang Q, Chen Y, Li T, Wang Q, et al. Identification and characterization of epithelial cells derived from human ovarian follicular fluid. Stem Cell Res Ther 2015;6:13.
26.	Lobb DK, Younglai EV. Ontario, Canada: A simplified method for preparing IVF granulosa cells for culture. J Assist Reprod Genet 2006;23:93.
27.	West-Farrell ER, Xu M, Gomberg MA, Chow YH, Woodruff TK, Shea LD. The mouse follicle microenvironment regulates antrum formation and steroid production: Alterations in gene expression profiles. Biol Reprod 2009;80:432-9.
28.	Liu C, Xia X, Miao W, Luan X, Sun L, Jin Y, et al. An ovarian cell microcapsule system simulating follicle structure for providing endogenous female hormones. Int J Pharm 2013;455:312-9.
29.	Kedem A, Fisch B, Garor R, Ben-Zaken A, Gizunterman T, Felz C, et al. Growth differentiating factor 9 (GDF9) and bone morphogenetic protein 15 both activate development of human primordial follicles in vitro, with seemingly more beneficial effects of GDF9. J Clin Endocrinol Metab 2011;96:E1246-54.
30.	Heise M, Koepsel R, Russell AJ, McGee EA. Calcium alginate microencapsulation of ovarian follicles impacts FSH delivery and follicle morphology. Reprod Biol Endocrinol 2005;3:47.
31.	Araújo VR, Gastal MO, Wischral A, Figueiredo JR, Gastal EL. In vitro development of bovine secondary follicles in two-and three-dimensional culture systems using vascular endothelial growth factor, insulin-like growth factor-1, and growth hormone. Theriogenology 2014;82:1246-53.
32.	Jonitz A, Lochner K, Peters K, Salamon A, Pasold J, Mueller-Hilke B, et al. Differentiation capacity of human chondrocytes embedded in alginate matrix. Connect Tissue Res 2011;52:503-11.
33.	Rao X, Huang X, Zhou Z, Lin XJ. An improvement of the 2ˆ (–delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath 2013;3:71.
34.	Yu X, Wang N, Ma Y, Wan Q, Qin M, Wang H. Generation of human oocyte-like cell differentiation in vivo. Sheng Wu Gong Cheng Xue Bao 2015;31:394-402.
35.	Vigo D, Villani S, Faustini M, Accorsi P, Galeati G, Spinaci M, et al. Follicle-like model by granulosa cell encapsulation in a barium alginate–protamine membrane. Tissue Eng 2005;11:709-14.
36.	Yin H, Kristensen SG, Jiang H, Rasmussen A, Andersen CY. Survival and growth of isolated pre-antral follicles from human ovarian medulla tissue during long-term 3D culture. Hum Reprod 2016;31:1531-9.
37.	Kreeger PK, Deck JW, Woodruff TK, Shea LD. The in vitro regulation of ovarian follicle development using alginate-extracellular matrix gels. Biomaterials 2006;27:714-23.
38.	Wei J, Han J, Zhao Y, Cui Y, Wang B, Xiao Z, et al. The importance of three-dimensional scaffold structure on stemness maintenance of mouse embryonic stem cells. Biomaterials 2014;35:7724-33.
39.	Canosa S, Adriaenssens T, Coucke W, Dalmasso P, Revelli A, Benedetto C, et al. Zona pellucida gene mRNA expression in human oocytes is related to oocyte maturity, zona inner layer retardance and fertilization competence. Mol Hum Reprod 2017;23:292-303.

Advanced Biomedical Research