RETRACTED ARTICLE: Production of viable chicken by allogeneic transplantation of primordial germ cells induced from somatic cells

Ruihua Zhao ,Jingyi Jiang,Qisheng Zuo,Zhenhua Zhao,Xiang Shi,Tingting Li,Xia Yuan,Jiancheng Li,Yang Zhang,Ming Zhang,Guobin Chang,Hongyan Sun,Qi Xu,Xinsheng Wu,Yani Zhang,Chen Zhang,Bing Li,Jing Jin,Ying Ding,Jiuzhou Song

doi:10.1038/s41467-021-23242-5

Abstract

The allogeneic transplantation of primordial germ cells (PGCs) derived from somatic cells overcomes the limitation of avian cloning. Here, we transdifferentiate chicken embryo fibroblasts (CEFs) from black feathered Langshan chickens to PGCs and transplant them into White Plymouth Rock chicken embryos to produce viable offspring with characteristics inherited from the donor. We express Oct4/Sox2/Nanog/Lin28A (OSNL) to reprogram CEFs to induced pluripotent stem cells (iPSCs), which are further induced to differentiate into PGCs by BMP4/BMP8b/EGF. DNA demethylation, histone acetylation and glycolytic activation elevate the iPSC induction efficiency, while histone acetylation and glycolytic inhibition facilitate PGCs formation. The induced PGCs (iPGCs) are transplanted into the recipients, which are self-crossed to produce 189/509 somatic cells derived chicken with the donor’s characteristics. Microsatellite analysis and genome sequencing confirm the inheritance of genetic information from the donor. Thus, we demonstrate the feasibility of avian cloning from somatic cells.

Highlights

The allogeneic transplantation of primordial germ cells (PGCs) derived from somatic cells overcomes the limitation of avian cloning
PGCs were induced by B27, epidermal growth factor (EGF), and basic fibroblast growth factor with an efficiency of 7%11, which was increased to 61.9% by addition of fetuin, retinoic acid (RA), bone morphogenetic protein 4 (BMP4), and activin A12
Our study demonstrates the possibility of chicken reproduction from somatic cells

Summary

Introduction

The allogeneic transplantation of primordial germ cells (PGCs) derived from somatic cells overcomes the limitation of avian cloning. We express Oct4/Sox2/Nanog/Lin28A (OSNL) to reprogram CEFs to induced pluripotent stem cells (iPSCs), which are further induced to differentiate into PGCs by BMP4/BMP8b/EGF. The induced PGCs (iPGCs) are transplanted into the recipients, which are self-crossed to produce 189/509 somatic cells derived chicken with the donor’s characteristics. Allogeneic transplantation of chicken primordial germ cells (PGCs) was shown to produce offspring[5] and may be used for cloning methods in avian species. Embryonic stem cells (ESCs) were induced to differentiate into PGCs when cocultured with BMP4-expressing trophoblasts in mice (2.9%) and became functional sperm. Induction of PGCs from somatic cells and the production of cloned offspring have never been achieved in chickens. No offspring were reported to be produced after in vivo transplantation

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