Leptin treatment of in vitro cultured embryos increases outgrowth rate of inner cell mass during embryonic stem cell derivation

In Vitro Cell Dev Biol Anim. 2019 Aug;55(7):473-481. doi: 10.1007/s11626-019-00367-y. Epub 2019 Jun 18.

Abstract

Leptin, a metabolic hormone, regulates the reproductive functions responding to both nutritional and body conditions. Embryonic stem cells play important roles in reproductive technology, but their derivation can be challenging. In this study, we evaluated the derivation rates of mouse embryonic stem cell (mESC) line from blastocysts developing in embryo culture media supplemented with different leptin concentrations. The results showed that addition of leptin into the embryo culture medium supported the in vitro development of mouse embryo. The mESC line derivation rates for media treated with 0, 10, 50, and 100 ng/ml of leptin were 61.24 % (54/88), 84.96 % (42/50), 81.79 % (61/76), and 85.78 % (56/67), respectively. In addition, leptin treatment of blastocysts upregulated the expression levels of the trophectoderm marker Cdx2, whereas inner cell mass markers Oct-4 and Nanog were not affected. mESC lines derived after leptin treatment demonstrated hallmarks of pluripotency, such as alkaline phosphatase activity, expression of, OCT4, NANOG, and SSEA1, as well as the ability to form embryoid bodies and well-differentiated teratomas. In conclusion, leptin has a positive effect on the derivation rate of mouse embryonic stem cell lines which may be, in part, due to its effects on the development of the trophectoderm cell lineage in the embryo.

Keywords: Development; Embryo; Leptin; Mouse; Stem cell.

MeSH terms

  • Animals
  • Blastocyst / cytology*
  • CDX2 Transcription Factor / biosynthesis
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Lineage
  • Cell Proliferation / drug effects*
  • Culture Media / pharmacology
  • Embryo Culture Techniques
  • Embryoid Bodies / cytology
  • Leptin / pharmacology*
  • Lewis X Antigen / biosynthesis
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mouse Embryonic Stem Cells / cytology*
  • Nanog Homeobox Protein / biosynthesis
  • Octamer Transcription Factor-3 / biosynthesis
  • Teratoma / chemically induced
  • Teratoma / metabolism*

Substances

  • CDX2 Transcription Factor
  • Cdx2 protein, mouse
  • Culture Media
  • Leptin
  • Lewis X Antigen
  • Nanog Homeobox Protein
  • Nanog protein, mouse
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse