The Effects Of Rabbit Allogeneic Serum And Tissue Lysates On Expansion And Differentiation Of MSCs

Mesenchymal stem cells( MSCs) are a kind of adult stem cell with self-renew and multilineage differentiation potential. Primarily owing to their low immunogencity, they will be wildly used for cell therapy and tissue engineering in the future. Finding suitable replacement for fetal bovine serum to culture human MSCs and avoid the risk from animals contamination is a hot spot today. In this paper, we have established a rabbit model to use allogeneic serum for the culture of MSCs.On the first part of the experiment, we have successfully separated and cultured the MSCs from rabbit marrow. We have got enough marrow from suitable sites and ficoll density gradient separation and whole -marrow culture method are used in our research. The prime cultures usually last 15 days. The cells then can be subcultured untill the 20th passage. The self-renew abilities of MSCs from new born rabbit and adult rabbit are compared and the cells from new born rabbit show greater expansion speed. The MSCs can be induced into osteoblasts and neural-like cells under certain conditions.At the second part of the experiment, we mixed serum from different rabbits. The Ig G is separated by ammonium sulfate precipitation and the complements are heat inactived. MSCs are cultured by these serum from prime passage to the third passage and induced into osteoblasts successfully. The self-renew abilities are also compared between cells cultured under fetal bovine serum, autologous serum and rabbit serum. We found the cells cultured under autologous serum expanded more rapidly than others. The difference between fetal bovine serum and rabbit serum are not show. Nature rabbit serum with Ig G and active complements do not support cell expansion .At the third part of the experiment, we studied many kinds of rabbit tissue (liver, brain, heart, kidney, spleen and thymus ) lysates and their function on the expansion and differentiation of MSCs. Liver lysates support the expansion of MSCs and brain lysates can induce the cells differentiate into neural-like cells.At the review part, this paper describes the development of the culture forMSCs during recent years. In this paper, we list many factors, including the age of the donors, the different solutions for isolation, the density for cell culture, the mediums and the surface materials for culture, and their effect on the cell expansion . Here we emphasize the development of different substitution, such as growth factors and human autologous serum, for fetal bovine serum (FBS) to avoid the risk from potential by virus. MSCs infected plasmids with telomerase gene have shown stronger expansion ability and multilineage differentiation potential. The development of perfusion culture systems will make the preparation of MSCs more suitable for clinical application.