Derivation of Mouse Parthenogenetic Advanced Stem Cells.

Mengyi Wei,Siqin Bao,Jia Liu,Xiaojie Yan,Chaoyue Zhao,Baojiang Wu,Shuo Cao,Jindun Zhang

doi:10.3390/ijms22168976

Mengyi Wei, Siqin Bao + Show 6 more

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https://doi.org/10.3390/ijms22168976

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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.

Highlights

epiblast stem cells (EpiSCs) have been obtained in activated activin A and FGF signaling in vitro, but whether EpiSCs can be derived from parthenogenetic blastocysts is still unknown [3,12,13]
We attempted to establish mouse parthenogenetic diploid epiblast stem cells derived from parthenogenetic blastocysts using a chemically defined medium
The parthenogenetic pluripotency embryonic stem cells (ESCs) has been derived from blastocysts stage; parthenogenetic EpiSCs have only been generated post-implantation, though the culture conditions used for all paESCs and paEpiSCs included serum or feeder [7,9,26,27] and there have no reports on the conversion of paEpiSCs to paESCs

Summary

Introduction

As parthenogenetic embryos can be obtained in vitro through incubation with 7% ethanol [3]. With the expression of imprinting genes, research on genetically edited parthenogenetic embryos and monogenetic individuals has begun to emerge [4,5,6]. In 2004, Tomohiro Kono produced adult parthenogenetic mice through the fusion of fully growing oocytes and non-growing oocytes with the deletion of the H19 gene region [4]. In 2008, Lee obtained parthenogenetic diploid stem cells from oocytes of preantral follicles using feeder, serum, and knockout serum replacement, which was able to differentiate into embryoid bodies in vitro and teratomas in vivo [7]. Qi Zhou exhibited the generation of both bimaternal and bipaternal mice from hypomethylated haploid ESCs carrying specific imprinted region deletions, but the live bipaternal mice shortly died after birth [8]

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