Reversine promotes porcine muscle derived stem cells (PMDSCs) differentiation into female germ-like cells

J Cell Biochem. 2012 Dec;113(12):3629-42. doi: 10.1002/jcb.24296.

Abstract

A small molecular chemical-Reversine has been shown to promote cell reprogramming and induce dedifferentiation of multiple terminally differentiated mesodermal origin cells, and then differentiate into other cell types within mesodermal lineages as well as neuroectodermal. However, the possibilities of these cells to give rise to germ cell lineages have not been examined. The objective of the current study was to detect the effect of Reversine on PMDSCs differentiation into germ cells. PMDSCs from fetal porcine skeletal muscle and their potential of differentiation into germ cells in vitro were investigated. The phenotype, proliferation potential, characteristic markers of the first adhesion cells (pp1), and the purified 2 times cells (pp3) were analyzed by growth curve, FACS, and RT-PCR, respectively. Then, the purified cells were induced with 10% or 20% bovine follicular fluid (FF), the results showed that some of the induced pp3 cells were similar as porcine oocyte, and expressed germ cell and oocyte markers analyzed by semi-quantitative RT-PCR and immunofluorescent staining. Reversine clearly increased the potentiality of PMDSCs differentiation into large round germ-like cells in FF induction medium analyzed by morpholgogy, QRT-PCR and immunofluoresce. The BrdU labeled PMDSCs might differentiate into female germ-like cells in recipient's kidney capsule, which were positive for germ cell and meiotic markers (Dazl, Vasa, Figla, Stra8, Scp3) and oocyte markers (Zp2, Zp3). These findings provided an efficient model to study the mechanism of cell proliferation and germ cell differentiation in livestock promoted by Reversine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Bromodeoxyuridine
  • Cattle
  • Cell Adhesion
  • Cell Differentiation / drug effects*
  • Cell Lineage
  • Cell Proliferation
  • Cell Shape
  • Culture Media / metabolism
  • Female
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Follicular Fluid / metabolism*
  • Meiosis
  • Mice
  • Morpholines / pharmacology*
  • Oocytes / cytology*
  • Oocytes / metabolism
  • Ovary / cytology
  • Phenotype
  • Purines / pharmacology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Satellite Cells, Skeletal Muscle / cytology*
  • Satellite Cells, Skeletal Muscle / drug effects
  • Stem Cell Transplantation
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • Swine

Substances

  • Biomarkers
  • Culture Media
  • Morpholines
  • Purines
  • Bromodeoxyuridine
  • 2-(4-morpholinoanilino)-6-cyclohexylaminopurine