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Production of Cloned-like Chicken by Allogeneic Transplantation of Somatic Cells Induced Primordial Germ Cells

ABSTRACT: Generation of genetically uniformed individuals from somatic cells is an effective approach for large-scale reproduction of elite varieties and a powerful tool for restoration of endangered species. However, this technique has never been realized in avian due to their oviparous reproduction pattern. In this study, we produced cloned-like chicken by allogeneic transplantation of somatic cells induced primordial germ cells (PGCs). Oct4, Sox2, Nanog and Lin28A (OSNL) factors were employed to reprogram primary chicken embryo fibroblasts (CEF) to pluripotent stem cells (iPS), in which DNA demethylation and histone acetylation were found to increase the efficiency to 13.00%. The obtained iPS presented embryonic stem cell like characters and were further induced to PGCs by BMP4/BMP8b/EGF, in which histone acetylation and glycolysis inhibition elevated the induction rate to 17.30% and 16.41%, respectively. With the optimized system, we induced Black Langshan Chicken (Gallus domestiaus, black feather) donated CEF to PGCs and transplanted them into the Recessive White Chicken (white feather) embryos. The transplanted cells migrated into the genital ridges and produced functional sperm or oocytes. The sexual matured recipients were self-crossed, with 189/509 (37.13%) cloned-like chicken produced. Microsatellite analysis confirmed their DNA inheritance from the black donor chicken. Thus, we demonstrated, for the first time, the feasibility of avian cloning from somatic cells.

ORGANISM(S): Gallus gallus

PROVIDER: GSE159511 | GEO | 2021/03/16

REPOSITORIES: GEO

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Project description:Eyal-Giladi and Kochav stage X (EGK.X) blastoderm mostly consists undifferentiated progenitor (pluripotent) cells that have the potency to differentiate into all cell types. Analyzing the transcriptional programs of blastoderm is critical to understand the molecular mechanisms that underlie the developmental fate of early embryos, and practical application of blastoderm-derived pluripotent cells, including ESC. Primordial germ cells (PGC) are also undifferentiated progenitor cells that later differentiate into male and female germ cells. Analyzing the transcriptional programs of PGC is critical to understand the molecular mechanisms that underlie the developmental fate of early germ cells, and practical application of PGC in generating transgenic chickens. Therefore, we performed whole-transcriptome sequencing (WTS) in the chicken EGK.X blastoderm and PGCs together with the chicken embryonic fibroblasts (CEF, as a reference sample). CEF are differentiated and an abundant somatic cells. The generated WTS data of CEF, EGK.X blastoderm and PGC are rich resources to utilize for various focused research. As a first focused research, we utilized the generated WTS data for analyzing the expression profiling of DNA repair and apoptosis pathway genes in EGK.X blastoderm and PGC compared with CEF. DNA is the fundamental unit of inheritance and is susceptible to damage by various exogenous and endogenous sources. When the DNA is damaged, the cell sense the nature of damage through multiple surveillance mechanisms and select an appropriate DNA repair pathway to repair the damage and avoid passing the damage to a progeny of cells. When the cell fails to repair DNA damage, cell cycle progression is halted and apoptosis is initiated. Despite several genes are involved in five major DNA repair pathways and two major apoptosis pathways, a comprehensive understanding on the expression of those genes are not well-understood in chicken tissues. Through WTS analysis in CEF, stage X blastoderm and PGC, we identified that most of the DNA repair pathway genes were expressed more high in blastoderm and high in PGC than CEF. At the other hand, most of the apoptosis pathway genes were expressed low in blastoderm and PGC than CEF.

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Production of Cloned-like Chicken by Allogeneic Transplantation of Somatic Cells Induced Primordial Germ Cells

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