Construction of human cell culture system for regenerative medicine without using animal components

点击翻译按钮获取中文摘要

It is very important to expand the number of human hematopoietie stem cells in cord blood without differentiation, because the cell number is not enough for patients of heavy weight. In addition, in the case human embryonic stem cell there is no way to culture without murine fibroblast cells. Therefore, we have tried to develop culture systems of human stem cells without using animal feeder cells.1.New human bone marrow cells incorporated with telomerase gene were established with cooperation with Cancer Research Center. Some cell lines were cloned and the clones supported growth of hematopoietie stem cells(HSC) in cord blood. However, in these cases, the growth of cell line was too fast to isolate HSC from the cell line. Therefore, the cell line was chemically fixed and employed for supporting growth of HSC.2.When some cells supporting growth of HSC were induced to fat cells, the supporting abilities disappeared. Therefore, the difference of proteins before and after the differentiation was investigated and some differences were observed by two dimensional electrophoresis. Now the gene expression is also investigated by a DNA microarray. When the protein or gene for supporting growth ofHSC, the protein will be immobilized on a substrate for artificial cell culture matrix.3.Monkey embryonic stem(ES) cell was cultured on human placenta feeder layers without differentiation of ES cells. Considering that he placenta was usually discarded as a medical waste, the success is very useful. Although the monkey ES cell was used as a preclinical research, the similarity can be extended to human ES cells and it will be very useful for practical regenerative medicine.4.Mouse ES cells were cultured on various artificial substrata and the growth was evaluated. It was found that chemically fixed feeder cell supported the growth of murine ES cells. Now this fixation is applied for placenta for culture of monkey ES cells.