Heterogeneity Of Donor Somatic Cells And Its Effect On The Development Efficiency Of Porcine Cloned Embryos

Porcine somatic cell cloning technology,also known as somatic cell nuclear transfer technique,has very important application value in the fields of agricultural production,disease animal model preparation,reprogramming mechanism research and human biomedical science and other fields.However,the efficiency of porcine cloned embryos to birth is only about 1%,which seriously restricts the development and application of porcine somatic cell nuclear transfer technology.The developmental efficiency of cloned embryos depends on whether the gene expression pattern is correct in the development process.Studies have shown that even though there are significant differences in gene expression patterns in different individual somatic cells of the same source,this difference may result in differences in gene expression patterns in the development of cloned embryos produced by different mononuclear cell nuclear transplants from the same source,leading to differences in their developmental efficiency.The efficiency of porcine cloning will be significantly enhanced if the "elite" donor cells for nuclear transplant embryos can be screened from the somatic cells of the same source for nuclear transplantation.In this study,porcine fibroblasts from the same source were cultured by single cell inoculation to obtain a group of monoclonal clones,each single cell clone randomly picks 1 or 2 single cells for transcription sequencing,and selects the gene expression pattern ‘alternative’ single cell clone and gene expression pattern ‘normal’ according to the sequencing results of single cell transcription group.The differences in in vitro development efficiency of two cloned embryos prepared by ‘alternative’ and ‘ordinary’ single cell clones were compared by somatic cell nuclear transfer.The experimental results show that(1)In the detection of two single cell transcriptome of 11 single cell clones,only 3 single cell clones of two Single-cell transcription groups were not similar,while 8 single cell clones of two Single-cell transcription groups were similar,proving that most of the cases originated from the same single cell clones of different single-cell gene expression patterns similar.(2)Of the 52 single cell of the same source,there are 48 single cell transcriptome is similar,but 4 Single-cell(numbered D11-1,D12-1,DW61-2,DW99-2)transcription groups were not similar to 48 ‘normal’ Single-cell transcription groups,and the 4 ‘alternative’ Single-cell transcription groups were not identical,The implication is that the 4 ‘alternative’ single-celled cells may contain ‘elite’ cell that facilitates the reprogramming of nuclear transplants.(3)There was no significant difference in the in vitro development efficiency between the two groups of porcine cloned embryos which were derived from ‘alternative’ cloned cells D11 and ‘normal’ cloned cells D28.This study provides a basis for the establishment of a new method based on screening donor cells to improve the efficiency of porcine somatic cell cloning,which provides a basis for revealing the molecular mechanism of epigenetic reprogramming in porcine somatic cells during nuclear transplantation.