title: Overexpression of CD61 promotes hUC-MSC differentiation into male germ-like cells.
abstract: OBJECTIVE
  Previous studies have shown that germ-like cells can be induced from human umbilical cord mesenchymal stem cell (hUC-MSCs) in vitro. However, induction efficiency was low and a stable system had not been built. CD61, also called integrin-β3, plays a significant role in cell differentiation, in that CD61-positive-cell-derived pluripotent stem cells easily differentiate into primordial germ-like cells (PGC). Here, we have explored whether overexpression of CD61 would promote hUC-MSC differentiation into PGC and male germ-like cells.
  
  
  METHODS
  hUC-MSCs were cultured and transduced using pCD61-CAGG-TRIP-pur (oCD61) and pTRIP-CAGG plasmid (Control), and hUC-MSCs overexpressed CD61 were induced by bone morphogenetic protein 4 (BMP4, 12.5 ng/ml), to differentiate into PGC and male germ cells. Quantitative real-time PCR (RT-qPCR), western blotting and immunofluorescence staining were used to examine PGC- and germ cell-specific markers.
  
  
  RESULTS
  High expression levels of PGC-specific markers were detected in oCD61 hUC-MSCs compared to controls. After BMP4 induction, expression levels of male germ cell markers such as Acrosin (ACR), Prm1 and meiotic markers including Stra8, Scp3 in oCD61 were significantly higher than those of the Control group.
  
  
  CONCLUSIONS
  Under induction of BMP4, CD61-overexpressing hUC-MSCs, which had turned into PGC-like cells, could be further differentiated into male germ-like cells. Thus, a simple and efficient approach to study male germ cell development by using hUC-MSCs has been established.
authors: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors: Bo
authors: B
authors: Li
authors: Department of Pathology, Yangling Demonstration Zone Hospital, Yangling, Shaanxi, 712100, China.
authors: Weishuai
authors: W
authors: Liu
authors: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors: Mengru
authors: M
authors: Zhuang
authors: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors: Na
authors: N
authors: Li
authors: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors: Siyu
authors: S
authors: Wu
authors: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors: Shaohui
authors: S
authors: Pan
authors: College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
authors: Jinlian
authors: J
authors: Hua
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002454:Cell Differentiation; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D005854:Germ Cells / Q000166:cytology*; D006801:Humans; D039661:Integrin beta3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008954:Models, Biological; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D020539:Sequence Analysis, Protein; D012689:Sequence Homology, Nucleic Acid; D014470:Umbilical Cord / Q000166:cytology*
chemical_list: D015415:Biomarkers; D055415:Bone Morphogenetic Protein 4; D003062:Codon; D039661:Integrin beta3