Screening of serum-free culture system for umbilical cord mesenchymal stem cells

ObjectiveTo screen serum-free medium for culturing umbilical cord mesenchymal stem cells.

MethodsNeonatal umbilical cord specimens were collected and cut into tissue fragments.The samples were randomly divided into group A, group B and group C, and were cultured with three different serum-free media MesenCult, Ultraculture and AM-V, respectively.The migration time and cell morphology of the primary umbilical cord cells in the three groups were observed, and the proliferation ability, differentiation ability and surface antigen expression of the cells were detected.

ResultsThe average time for primary cells to grow out from the tissue blocks in group A, group B and group C was 17 d, 6 d and 7 d.Observed under the microscope, the cells growing out of the tissue blocks were scattered distributed, with short spindle or polygonal shape, full shape and good refraction.When growing to fusion, the cells in group A and group B displayed spiral growth, with good refraction and similar size, but cells in group C were slender, slightly worse in refraction, and had no obvious feature of spiral growth.Cells in the three groups all expressed CD105, CD90, CD73 antigens, but did not express CD34, CD45, HLA-DR antigens, and there was no significant difference in the expression of the above markers among the three groups(P>0.05).Mesenchymal stem cells in the three groups all had the ability to differentiate into adipocytes and osteoblasts, and the differentiation ability in group C was weaker than that in group A and group B.

ConclusionsThe culture system in group B has a shorter time, better stem cell characteristics and reasonable price, which can be used as the first choice medium for large-scale culture of human mesenchymal stem cells.