Induction differentiation of equine BMSCs into chondrocytes and screening analysis of key IncRNAs-mRNAs
In order to further screen for the key IncRNAs and mRNAs of bone marrow mesenchymal stem cells induced differentiation into chondrocytes,equine BMSCs were selected as the research object,and whole transcriptome sequencing of cells were carried out at three different induced differentiation stages into chondrocyte.And then lncRNAs and mRNAs with important functions in the differentiation of BMSCs into chondrocytes were screened out.At the same time,mRNAs,lncRNAs and their target genes were verified,and then the regulatory network of lncRNAs in the differentiation of BMSCs into chondrocytes were constructed.The results showed that:1)Equine BMSCs line was established in vitro and the cells were successfully induced and differentiated into chondrocytes.2)A total of 3 592 lncRNAs were identified by high-throughput transcriptome sequencing of BMSCs at different differentiation stages,including 250 differentially high expression lncRNAs and 116 differentially low expression lncRNAs,280 differentially high expression mRNAs,and 105 differentially low expression mRNAs.3)Five potential candidate factors regulating the differentiation of horse BMSCs into chondrocytes were screened and validated,including lnc-cpm,lnc-traf3,lnc-22173,lnc-acvr2a,lnc-pde4d,as well as nine corresponding target genes MDM2,TRAF3,USP25,ACVR2A,PDE4D,as well as BCL2,WAF1,SIRT1,NRF2.4)A differentially expressed lncRNA-mRNA interaction regulatory network was constructed,which was mainly enriched in following signaling pathway,including Wnt,p53,MAPK and NF-κb.To sum up,in this study,differential genes were successfully screened out during the equine BMSCs into chondrocytes through whole transcriptome sequencing and analysis,which will lay a necessary foundation for further revealing the interaction mechanism between lncRNAs and mRNAs during chondrocyte differentiation,and provide a scientific basis for BMSCs been used in cell therapy for cartilage repair.