Cryopreserved Human Oocytes and Cord Blood Cells Can Produce Somatic Cell Nuclear Transfer-Derived Pluripotent Stem Cells with a Homozygous HLA Type.

Jeoung Eun Lee,Soo Kyung Jung,Jin Hee Eum,Dong-Won Seol,Kyungsoo Ha,Chang-Hwan Park,Tae Ki Yoon,Dong Ryul Lee,Jin Saem Lee,Hyun Soo Shin,Taehoon Chun,A-Reum Han,Ji Yoon Lee,Deok Rim Heo,Jung Ho Im

doi:10.1016/j.stemcr.2020.05.005

Abstract

SummaryHuman pluripotent stem cells (PSCs) through somatic cell nuclear transfer (SCNT) may be an important source for regenerative medicine. The low derivation efficiency of stem cells and the accessibility of human oocytes are the main obstacles to their application. We previously reported that the efficiency of SCNT was increased by overexpression of H3K9me3 demethylase. Here, we applied a modified derivation method to the PSC line and first obtained human SCNT-PSC lines derived from both donated cryopreserved oocytes and cord blood cells with a homozygous human leukocyte antigen (HLA) type. The SCNT-PSCs have very similar characteristics with embryonic stem cells (ESCs) and additionally have shown immunocompatibility in an in vitro and in vivo humanized mouse with a matching HLA type. Our study demonstrates that SCNT technology using donated cryopreserved oocytes and cord blood cells with a known HLA type provides a promising method for establishing a human HLA-matched SCNT-PSC bank for regenerative medicine.

Highlights

It appears that a major hurdle for the application of human Somatic cell nuclear transfer (SCNT)-pluripotent stem cells (PSCs) as sources for cell therapy has been partially overcome, further research is still needed for better embryonic development
Derivation of Human SCNT-PSCs Using Cryopreserved Human Oocytes and Its Characterization To analyze the potential of cryopreserved human oocyte cytoplasm for SCNT-reprogramming, two types of nuclear donor cells were prepared for SCNT
One of them was developed further to an expanded blastocyst at 6 days (Figure 1B). This expanded blastocyst was plated onto irradiated mouse embryonic fibroblast (MEF) cells and eventually resulted in a stable human SCNT-PSC line (CHA-FT-NT17, Figure 1B)

Summary

Introduction

Somatic cell nuclear transfer (SCNT) mediated by oocytes can produce cloned embryos in more than 20 species (Gurdon, 1962; Kato et al, 1998; Rodriguez-Osorio et al, 2012; Wakayama et al, 1998; Wilmut et al, 1997), and it has contributed to the establishment of pluripotent stem cells (PSCs) in various mammals, including humans (Chung et al, 2014; Tachibana et al, 2013; Yamada et al, 2014). The number of cloned eggs arrested at the eightcell stage was greatly reduced, and the SCNT-PSC derivation rate increased to 7.1% per donated mature oocyte (Chung et al, 2015) This technology has recently been applied to non-human primate cloning systems and successfully produced cloned macaque monkeys (Liu et al, 2018). It appears that a major hurdle for the application of human SCNT-PSCs as sources for cell therapy has been partially overcome, further research is still needed for better embryonic development

Results

Discussion

Conclusion