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Inhibiting transforming growth factor‐β signaling regulates in vitro maintenance and differentiation of bovine bone marrow mesenchymal stem cells
Author(s) -
Zhao XinXin,
An XingLan,
Zhu XianChun,
Jiang Yu,
Zhai YanHui,
Zhang Sheng,
Cai NingNing,
Tang Bo,
Li ZiYi,
Zhang XueMing
Publication year - 2018
Publication title -
journal of experimental zoology part b: molecular and developmental evolution
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.823
H-Index - 63
eISSN - 1552-5015
pISSN - 1552-5007
DOI - 10.1002/jez.b.22836
Subject(s) - microbiology and biotechnology , homeobox protein nanog , cd90 , stem cell , cellular differentiation , biology , mesenchymal stem cell , sox2 , chemistry , cd34 , embryonic stem cell , induced pluripotent stem cell , biochemistry , gene
Bovine bone marrow mesenchymal stem cells (bBMSC) are potential stem cell source which can be used for multipurpose. However, their application is limited because the in vitro maintenance of these cells is usually accompanied by aging and multipotency losing. Considering transforming growth factor‐β (TGF‐β) pathway inhibitor Repsox is beneficial for cell reprogramming, here we investigated its impacts on the maintenance and differentiation of bBMSC. The bBMSC were enriched and characterized by morphology, immunofluorescent staining, flow cytometry, and multilineage differentiation. The impacts of Repsox on their proliferation, apoptosis, cell cycle, multipotency, and differentiation were examined by Cell Counting Kit‐8 (CCK‐8), real‐time polymerase chain reaction, induced differentiation and specific staining. The results showed that highly purified cluster of diffrentiation 73 + (CD73 + )/CD90 + /CD105 + /CD34 − /CD45 − bBMSC with adipogenic, osteogenic, and chondrogenic differentiation capacities were enriched. Repsox treatments (5 μM, 48 hr) enhanced the messenger RNA mRNA levels of the proliferation gene (telomerase reverse transcriptase [ TERT ]; basic fibroblast growth factor [ bFGF ]), apoptosis‐related gene ( bax and Bcl2 ), antiapoptosis ratio ( Bcl2/bax ), and pluripotency marker gene ( Oct4, Sox2, and Nanog ), instead of changing the cell cycle, in bBMSC. Repsox treatments also enhanced the osteogenic differentiation but attenuated the chondrogenic differentiation of bBMSC, concomitant with decreased Smad2 and increased Smad3/4 expressions in TGF‐β pathway. Collectively, inhibiting TGF‐β/Smad signaling by Repsox regulates the in vitro maintenance and differentiation of bBMSC.