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BMP4 is insufficient to differentiate umbilical cord mesenchymal stem cells into germ cell-like cells in vitro
, 1, 1
Affiliations- Xinxiang Medical University, Xinxiang, China
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Abstract
Objectives: Mesenchymal stem cell (MSC)-based therapies are expected to restore the fertility of infertile patients. In addition to MSC-derived paracrine effects to improve reproductive function, the differentiation of MSCs into germ cell (GC)-like cells is still a promising method to repair the injured reproductive system. The aim of this study was to examine the effect and potential mechanism of BMP4 in inducing umbilical cord MSC (UcMSC) transdifferentiation into GC-like cells. Material and methods: UcMSCs were isolated, cultured and identified by flow cytometry and multilineage differentiation assays. After induction with 12.5 ng/mL BMP4 for 21 days, UcMSCs were collected for further examination. Immunofluorescence was used to detect the expression of Prdm1 and Prdm14; RT-PCR and RNA sequencing were used to detect differential gene expression (DEGs). Results: The morphology of UcMSCs became large and flat after treatment with BMP4; the expression of GC-related genes (OCT4, Prdm1, Ifitm3 and Stella) was significantly downregulated, and further immunofluorescence results also confirmed the significant downregulation of Prdm1 in UcMSCs with BMP4 induction, while the expression of Prdm14 was significantly upregulated. The results of RNA sequencing and further analysis revealed no explicit correlation between BMP4 induction and the differentiation of UcMSCs into GC-like cells based on the 662 screened DEGs in UcMSCs with or without BMP4 induction. Conclusions: The differentiation of MSCs into GC-like cells is rather complex, and BMP4 alone is insufficient to induce UcMSCs to differentiate into GC-like cells, regardless of protein level or gene expression level.
Abstract
Objectives: Mesenchymal stem cell (MSC)-based therapies are expected to restore the fertility of infertile patients. In addition to MSC-derived paracrine effects to improve reproductive function, the differentiation of MSCs into germ cell (GC)-like cells is still a promising method to repair the injured reproductive system. The aim of this study was to examine the effect and potential mechanism of BMP4 in inducing umbilical cord MSC (UcMSC) transdifferentiation into GC-like cells. Material and methods: UcMSCs were isolated, cultured and identified by flow cytometry and multilineage differentiation assays. After induction with 12.5 ng/mL BMP4 for 21 days, UcMSCs were collected for further examination. Immunofluorescence was used to detect the expression of Prdm1 and Prdm14; RT-PCR and RNA sequencing were used to detect differential gene expression (DEGs). Results: The morphology of UcMSCs became large and flat after treatment with BMP4; the expression of GC-related genes (OCT4, Prdm1, Ifitm3 and Stella) was significantly downregulated, and further immunofluorescence results also confirmed the significant downregulation of Prdm1 in UcMSCs with BMP4 induction, while the expression of Prdm14 was significantly upregulated. The results of RNA sequencing and further analysis revealed no explicit correlation between BMP4 induction and the differentiation of UcMSCs into GC-like cells based on the 662 screened DEGs in UcMSCs with or without BMP4 induction. Conclusions: The differentiation of MSCs into GC-like cells is rather complex, and BMP4 alone is insufficient to induce UcMSCs to differentiate into GC-like cells, regardless of protein level or gene expression level.
Keywords
germ cell; mesenchymal stem cell; bone morphogenetic protein 4; fertility
About this articleTitle
BMP4 is insufficient to differentiate umbilical cord mesenchymal stem cells into germ cell-like cells in vitro
Journal
Issue
Article type
Research paper
Pages
64-72
Published online
2021-11-22
Page views
3560
Article views/downloads
746
DOI
Pubmed
Bibliographic record
Ginekol Pol 2023;94(1):64-72.
Keywords
germ cell
mesenchymal stem cell
bone morphogenetic protein 4
fertility
Authors
Pengbo Wang
Huanhuan Hu
Xinyi Li
Ruiyun Zhang
Hongbin Cheng
Haixia Qin
Guojie Ji
Huigen Feng
Yanli Liu
Juntang Lin
References (26)- Guo X, Tang Y, Zhang P, et al. Effect of ectopic high expression of transcription factor OCT4 on the "stemness" characteristics of human bone marrow-derived mesenchymal stromal cells. Stem Cell Res Ther. 2019; 10(1): 160.
- Asgari HR, Akbari M, Yazdekhasti H, et al. Comparison of human amniotic, chorionic, and umbilical cord multipotent mesenchymal stem cells regarding their capacity for differentiation toward female germ cells. Cell Reprogram. 2017; 19(1): 44–53.
- Samiec M, Opiela J, Lipiński D, et al. Trichostatin A-mediated epigenetic transformation of adult bone marrow-derived mesenchymal stem cells biases the in vitro developmental capability, quality, and pluripotency extent of porcine cloned embryos. Biomed Res Int. 2015; 2015: 814686.
- Ling Li, Feng X, Wei T, et al. Human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation improves ovarian function in rats with premature ovarian insufficiency (POI) at least partly through a paracrine mechanism. Stem Cell Res Ther. 2019; 10(1): 46.
- Zheng Q, Fu X, Jiang J, et al. Umbilical cord mesenchymal stem cell transplantation prevents chemotherapy-induced ovarian failure via the NGF/TrkA pathway in rats. Biomed Res Int. 2019; 2019: 6539294.
- Dessouky AA, Gouda ZA, Arafa MAA, et al. Hypoxia-preconditioned human umbilical cord blood-derived mesenchymal stem cells mitigate hypoglycemic testicular injury induced by insulin in rats. Cells Tissues Organs. 2020; 209(2-3): 83–100.
- Bochinski D, Lin GT, Nunes L, et al. The effect of neural embryonic stem cell therapy in a rat model of cavernosal nerve injury. BJU Int. 2004; 94(6): 904–909.
- Qiu X, Villalta J, Ferretti L, et al. Effects of intravenous injection of adipose-derived stem cells in a rat model of radiation therapy-induced erectile dysfunction. J Sex Med. 2012; 9(7): 1834–1841.
- Ohinata Y, Ohta H, Shigeta M, et al. A signaling principle for the specification of the germ cell lineage in mice. Cell. 2009; 137(3): 571–584.
- Wei Y, Fang J, Cai S, et al. Primordial germ cell-like cells derived from canine adipose mesenchymal stem cells. Cell Prolif. 2016; 49(4): 503–511.
- Ghorbanlou M, Abdanipour A, Shirazi R, et al. Indirect co-culture of testicular cells with bone marrow mesenchymal stem cells leads to male germ cell-specific gene expressions. Cell J. 2019; 20(4): 505–512.
- Kashyap V, Rezende NC, Scotland KB, et al. Regulation of stem cell pluripotency and differentiation involves a mutual regulatory circuit of the NANOG, OCT4, and SOX2 pluripotency transcription factors with polycomb repressive complexes and stem cell microRNAs. Stem Cells Dev. 2009; 18(7): 1093–1108.
- Huang P, Lin LiM, Wu XY, et al. Differentiation of human umbilical cord Wharton's jelly-derived mesenchymal stem cells into germ-like cells in vitro. J Cell Biochem. 2010; 109(4): 747–754.
- Lee YM, Kim TH, Lee JH, et al. Overexpression of Oct4 in porcine ovarian stem/stromal cells enhances differentiation of oocyte-like cells in vitro and ovarian follicular formation in vivo. J Ovarian Res. 2016; 9: 24.
- Li N, Pan S, Zhu H, et al. BMP4 promotes SSEA-1(+) hUC-MSC differentiation into male germ-like cells in vitro. Cell Prolif. 2014; 47(4): 299–309.
- Mazaheri Z, Movahedin M, Rahbarizadeh F, et al. Different doses of bone morphogenetic protein 4 promote the expression of early germ cell-specific gene in bone marrow mesenchymal stem cells. In Vitro Cell Dev Biol Anim. 2011; 47(8): 521–525.
- Liu Y, Niu R, Li W, et al. Biological characteristics of human menstrual blood-derived endometrial stem cells. J Cell Mol Med. 2018; 22(3): 1627–1639.
- Ding L, Yan G, Wang B, et al. Transplantation of UC-MSCs on collagen scaffold activates follicles in dormant ovaries of POF patients with long history of infertility. Sci China Life Sci. 2018; 61(12): 1554–1565.
- Lai D, Wang F, Yao X, et al. Human endometrial mesenchymal stem cells restore ovarian function through improving the renewal of germline stem cells in a mouse model of premature ovarian failure. J Transl Med. 2015; 13: 155.
- Mochizuki K, Tando Y, Sekinaka T, et al. SETDB1 is essential for mouse primordial germ cell fate determination by ensuring BMP signaling. Development. 2018; 145(23).
- Murakami K, Günesdogan U, Zylicz JJ, et al. NANOG alone induces germ cells in primed epiblast in vitro by activation of enhancers. Nature. 2016; 529(7586): 403–407.
- Shirazi R, Zarnani AH, Soleimani M, et al. Differentiation of bone marrow-derived stage-specific embryonic antigen 1 positive pluripotent stem cells into male germ cells. Microsc Res Tech. 2017; 80(4): 430–440.
- Katsara O, Mahaira LG, Iliopoulou EG, et al. Effects of donor age, gender, and in vitro cellular aging on the phenotypic, functional, and molecular characteristics of mouse bone marrow-derived mesenchymal stem cells. Stem Cells Dev. 2011; 20(9): 1549–1561.
- Ohinata Y, Payer B, O'Carroll D, et al. Blimp1 is a critical determinant of the germ cell lineage in mice. Nature. 2005; 436(7048): 207–213.
- Bao S, Leitch HG, Gillich A, et al. The germ cell determinant Blimp1 is not required for derivation of pluripotent stem cells. Cell Stem Cell. 2012; 11(1): 110–117.
- Wagner W, Horn P, Castoldi M, et al. Replicative senescence of mesenchymal stem cells: a continuous and organized process. PLoS One. 2008; 3(5): e2213.
