Derivation of Mouse Parthenogenetic Advanced Stem Cells

Int J Mol Sci. 2021 Aug 20;22(16):8976. doi: 10.3390/ijms22168976.

Abstract

Parthenogenetic embryos have been widely studied as an effective tool related to paternal and maternal imprinting genes and reproductive problems for a long time. In this study, we established a parthenogenetic epiblast-like stem cell line through culturing parthenogenetic diploid blastocysts in a chemically defined medium containing activin A and bFGF named paAFSCs. The paAFSCs expressed pluripotent marker genes and germ-layer-related genes, as well as being alkaline-phosphatase-positive, which is similar to epiblast stem cells (EpiSCs). We previously showed that advanced embryonic stem cells (ASCs) represent hypermethylated naive pluripotent embryonic stem cells (ESCs). Here, we converted paAFSCs to ASCs by replacing bFGF with bone morphogenetic protein 4 (BMP4), CHIR99021, and leukemia inhibitory factor (LIF) in a culture medium, and we obtained parthenogenetic advanced stem cells (paASCs). The paASCs showed similar morphology with ESCs and also displayed a stronger developmental potential than paAFSCs in vivo by producing chimaeras. Our study demonstrates that maternal genes could support parthenogenetic EpiSCs derived from blastocysts and also have the potential to convert primed state paAFSCs to naive state paASCs.

Keywords: epiblast stem cells; parthenogenetic diploid stem cells; parthenogenetic embryos; pluripotency.

MeSH terms

  • Activins / metabolism
  • Animals
  • Blastocyst / metabolism
  • Bone Morphogenetic Protein 4 / pharmacology
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects
  • DNA Methylation / drug effects
  • Embryo Culture Techniques / methods
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism*
  • Female
  • Fibroblast Growth Factors / pharmacology
  • Germ Layers / metabolism
  • Germ Layers / physiology
  • Leukemia Inhibitory Factor / pharmacology
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred ICR
  • Mouse Embryonic Stem Cells / cytology
  • Parthenogenesis / genetics
  • Parthenogenesis / physiology*
  • Pluripotent Stem Cells / metabolism
  • Pluripotent Stem Cells / pathology

Substances

  • Bone Morphogenetic Protein 4
  • Leukemia Inhibitory Factor
  • activin A
  • Activins
  • Fibroblast Growth Factors