Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

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Oct4 Methylation in Induced Differentiation of Bone Mesenchymal Stem Cells

WANG Ju1,FU Yu hao2,WANG Wei shan3,WANG Duan ming1,ZHOU Zong yao1   

  1. 1Department of Histology and Embryology,School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China
    2Hainan Provincial Center of Reproductive Medicine,Haikou 570102,China
    3Department of Orthopaedics,First Affiliated Hospital of Shihezi University,Shihezi,Xinjiang 832000,China
  • Online:2013-07-04 Published:2013-07-04

Abstract:

To investigate the methylation Oct4 in orientationinduced differentiation in bone marrow mesenchymal stem cells Methods Mice BMSCs were isolated and purified from bone marrow by adherent culture,and then identified by morphology and immunocytochemistryMouse osteoblastic cells were cultured by bone fragments inoculation,and then identified by alkaline phosphatase(AKP)staining and alizarin red stainingBMSCs were induced to differentiate into osteoblasts in vitro Indirect immunofluorescence staining and reverse transcription polymerase chain reaction(RTPCR)were used to detect the expressions of Oct4 in BMSCs before and after inductionThe methylation status of Oct4 gene in mouse BMSCs was explored by a methylationspecific PCR before and after induction Results The isolated mice BMSCs massively proliferated in vitro and formed cell colones with uniform morphologyPositive expressions of CD29,cKit,and CD44 and negative expression of CD34 were found in the isolated cellsAfter 10 days[DK]’[DK] induction,both AKP and the alizarin red were positive in cells and osteoblastic cells isolated from mice skull bonesThe indirect immunoinfluorescence staining and RTPCR also showed that the Oct4 expression in the directed differentiation of mouse BMSCs was downregulatedThe CpG island of Otc4 gene promoter in mouse BMSCs became methylated during the induced differentiation Conclusions Mice BMSCs and osteoblasts were successfully cultured in vitro in this studyOct4 may be involved in the maintenance of adult stem cell pluripotencyThe downregulated expression of Oct4 gene in mouse BMSCs during the directed differentiation may contribute to the methylation of CpG island in Otc4 gene promoter

Key words: Oct4, bone marrowderived mesenchymal stem cells, differentiation, methylation

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