Comparison Of Biological Characteristics Of CD146 Positive Subpopulations Of Mesenchymal Stem Cells Derived From Three Different Sources

Mesenchymal stem cells(MSCs)are a kind of pluripotent stem cells that have been studied extensively.They have the potential for multi-directional differentiation,self-renewal and possess low immunogenicity.They come from a wide range of origins,such as umbilical cord,adipose tissue,bone marrow,dental pulp and other tissues.MSCs have the ability of long-term adherent when culture in vitro,and can be induced to differentiate into osteoblasts,adipocytes,chondroblasts under different inducing conditions,and even can differentiate trans-germ layers.CD 146 molecule,also known as melanoma cell adhesion molecule(MCAM),is a member of the immunoglobulin superfamily.CD 146 molecule is mainly expressed on endothelial cells,which can affect endothelial cell proliferation,migration,and angiogenesis.It is an important molecule that affects vascular structure,vascular permeability,and angiogenesis.CD 146 molecule is also expressed on smooth muscle cells and pericytes.It can participate in various biological processes,including angiogenesis,tumor metastasis,lymphocyte activation,tissue development and regeneration.However,the biological characteristics and functions of 146 positive MSCs from different sources are not clear.In our study,we introduced magnetic activated cell sorting(MACS)to obtain highly purified CD146+subpopulations of mesenchymal stem cells from three different origins,umbilical cord,bone marrow,and adipose tissue.We then compared the morphology,proliferative ability,surface markers and biological characteristics such as the adipogenesis,osteogenesis,and angiogenesis capacity of MSCs from the three origins.The phenotype was analyzed by flow cytometry;the cell structure was observed by transmission electron microscopy;the proliferation ability was detected by MTS,cell cycle,etc.;The differentiation abilities were tested by adipogenic and osteogenic inducement and oil red O staining was used to detect adipogenic differentiation while ALP staining was applied to detect osteogenic differentiation.The expression levels of genes related to adipogenesis(LPL,C/EBPa and PPARy),osteogenesis(ALP,OPN and RUNX2),stemness(OCT-4,SOX2 and NANOG)and angiogenesis(bFGF,VEGF were all determined by RT-qPCR;Western blot was used to detect the protein expression of adipogenesis and osteogenesis;induction of angiogenesis in vitro test the ability of cells to form tubes in vitro.We found that the three types of cells were similar in morphology after sorting,but CD146+UC-MSCs have stronger proliferation capacity whereas CD146+BM-MSCs have more abundant mitochondria.Under the same inducing conditions,CD146+AD-MSCs have stronger adipogenic differentiation ability,and CD 146+BM-MSCs have stronger osteogenic differentiation ability,which may be related to its tissue origin.CD146+AD-MSCs expressed higher levels of three stemness genes,OCT-4,SOX2 and NANOG.It has been reported that CD 146 is also expressed on pericytes,which can maintain the stability of blood vessels and play an important role in the process of angiogenesis.By comparing in vitro angiogenetic ability and angiogenesis-related gene expression,we found that CD146+BM-MSCs have better tube-forming ability.In summary,with the progress of the research on MSCs,its application is becoming more and more extensive.The comparison of the biological characteristics of CD146+subpopulation of mesenchymal stem cells derived from umbilical cord,adipose tissue and bone marrow provides new ideas for the selection of tissue engineering seed cells and establishes the foundation for clinical treatment using MSCs.