Induction Of Bone Marrow Mesenchymal Stem Cells By Oriented Or Co-culture Method

Objective To study the multitude potency of differentiating into adipocytes or nerval cells by oriented induction of bone marrow mesenchymal stem cells. Human bone marrow mesenchymal stem cells were induced towards pancreatic islet progenitor cells by co-culture system. The value of bone marrow mesenchymal stem cells in the treatment of diabetes mellitus was investigated.Methods The bone marrow mesenchymal stem cells were gained from sternum bone marrow in children aged 5～10 (Exclude the disease of hematopoietic system, under the permission of the parents). The human mesenchymal stem cells were isolated by Percoll lymphocyte separation medium. The third passages of MSCs were identificated by CD44-antibody,CD45-antibody and CD34-antibody. The fourth passage MSCs were induced by HG-DMEM containing 10^-8mmol/L dexamethasome and 10ug/mL insulin to differentiate into adipocytes which could be stained by oil red. Neural progenitor cells which could be identified by nestin staining were obtained by bFGF pretreatment, and then they were induced to form nerval network by 1%DMSO.Neonatus pancreatic mesenchymal cells were obtained by collagenase enzyme digestion, which were proliferated by serum-free LG-DMEM medium containing bFGF. Pancreatic mesenchymal cells were inoculated into the insert, and then the insert was put into the transwell plate which undedayer was covered with bone marrow mesenchymal stem cells. Co-culture method was adopted to induce bone marrow mesenchymal cells to differentiate into pancreatic islet progenitor cells initially.To establish the co-culture system including the bone marrow mesenchymal stem cells and the neural stem cells in order to induce the neural stem cells to differentiate into neurons.Results Adipocyte and neural cells were gained by the induction of bone marrow mesenchymal stem cells in vitro. Neonatus pancreatic mesenchymal cells containing nestin and CK-19 positive pancreatic stem cells could be induced to form island-like organ in vitro. Co-culture method could be adopted to induce bone marrow mesenchyme stem cells to differentiate into PDX-1 immunological reaction masccline cells. The co-culture system had effect on differentiation of neural stem cells.Conclusion The human bone mesenchymal stem cells could be isolated by Percoll lymphocyte separation medium. They could differentiate into adipocytes or neural cells. Foetus pancreatic gland microenvironment had effect on the differentiation of bone marrow mesenchymal stem cells towards pancreatic islet progenitor cells. Some substance secreted by bone marrow stem cells had effect on the differentiation of neural stem cells.