The invention discloses a preparation method of
bone marrow mesenchymal stem cells, and relates to the field of
stem cell preparation. The method comprises the following steps: mixing marrow fluid with 0.9% normal
saline, carrying out
density gradient centrifugation, and collecting mesenchymal stem cells from the lower layer of
plasma; culturing leucoderma cells by using a
cell culture plate coated with a CD73
antibody; when the cells grow to 70-80%, digesting the cells with
stem cell mild digestive enzymes; continuously culturing the
bone marrow mesenchymal stem cells by using a
cell cultureplate coated with a CD90
antibody; and when the cells grow to 70-80%, digesting the cells by using the
stem cell mild digestive enzymes, and collecting high-purity
bone marrow mesenchymal stem cells.According to the invention, the high-purity mesenchymal stem cells are separated from the marrow fluid by combining
gradient centrifugation adherence with two rounds of
antibody forward screening, a large number of miscellaneous cells do not exist in the culture process, the cells combined with the antibody cannot be eluted into the cells, and the
cell function is not affected. The preparation method disclosed by the invention is safe, and the capability of purifying and amplifying the
bone marrow mesenchymal stem cells is strong.