| The biological functions of mesenchymal stem cells (MSCs) from different cellular sources are not identical, due to the different microenvironments. The source of human MSCs is limited, so the therapeutic potentialities of MSCs which derived from animals have got more researchers’ attention. Porcine mesenchymal stem cells (MSCs) can be easily obtained from pigs and similar to human MSCs, they are excellent pre-clinical material. The genetic similarity of inbred line pig individual is high, so their MSCs are more stable and conducive to establishing a reliable, standard evaluation system in vitro. In this study, to reveal the molecular mechanisms of the tissue repair potential of MSCs, we isolated bone marrow MSCs (BMMSCs) and umbilical cord MSCs (UCMSCs) from the Chinese inbred miniature pig-Wuzhishan pig (WZSP) by tissue anchored method, and indentified MSCs with detections of several surface antigens and osteogenesis, adipogenesis differentiation; generated genome-wide integrated maps of DNA methylation and mRNA transcripts by methylated DNA immunoprecipitation sequencing (MeDIP-Seq) and RNA-seq, analyzed biological function difference of MSCs which dependented on different tissue source. The results as follow:1. The adherent cells displayed a scatter distribution and grew in isolated clones. In BMMSCs, CD29, CD44and CD90were positively detected, whereas leucocyte antigen CD45and hematopoietic stem cell antigen CD34were not detected. In UCMSCs, CD90was detected, whereas the endothelial cell surface marker CD31, hematopoietic stem cell antigen CD34and leucocyte antigen CD45were not detected. Both BMMSCs and UCMSCs have the potential for osteogenic and adipogenic differentiation, and the osteogenic and adipogenie differentiation capacity of BMMSC were stronger than UCMSCs. These results illustrated that pure MSCs were obtained.2. Approximately147M reads (7.2Gb) of methylated DNA immunoprecipitation sequencing (MeDIP-seq) data were generated from each independent sample:BMMSCs and UCMSCs. The reads from the BMMSCs and UCMSCs samples aligned to75.52%and76.42%of the pig genome,58.98%of BMMSCs and00.89%of UCMSCs are the uniquely aligned non-duplicated reads, respectively. Nearly4.8Gb of RNA sequencing (RNA-seq) data were obtained from each samples, the reads from the BMMSC and UCMSC samples aligned to59.90%and59.83%of the pig genome,53.87%of BMMSCs and53.84%of UCMSCs are the uniquely aligned non-duplicated reads, respectively. The results of MeDIP-seq showed that:Genomic subtelomere regions of BMMSCs and UCMSCs exhibited a high methylation status. The methylation displayed in BMMSCs is higher than that of UCMSCs, indicating that the chromosomal structure of BMMSCs may be more stable. In two samples, both the promoter and genebody regions methylation were negatively correlated with the expression level of the genes, whereas the methylation of the promoter region displayed greater effects. Moreover, we obtained587genes displaying differences in promoter methylation,1979genes displaying expression differences, these genes were involved in many biological processes, declaring that the microenvironment difference of MSCs which dependented on different tissue source caused their biological function discrepancy. Further gene function analysis revealed that102genes displaying both expression and promoter methylation differences simultaneously, these genes were involved regulation of osteogenic, adipogenic, myocardial differentiation, cellular migration ability, and immunological competence, indicating that the regulatory role of DNA methylation of above genes, may be is one of the important reasons of make the tissue repair capacity difference between BMMSCs and UCMSCs.In conclusion, we have generated the first global integrated DNA methylation and transcriptional maps of porcine MSCs genomes, which illustrated that MSCs from different cellular sources could be used for different tissue repair engineering applications. This study provides a molecular theory basis for the application of human MSCs. |
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