Abstract:

BACKGROUND: It is important to search a rational cell implantation density to achieve stably and high efficiently growing stem cells during the culture of stem cells.
OBJECTIVE: To investigate the effects of different implantation densities on primary culture of human umbilical cord mesenchymal stem cells and their differentiation into neuron-, islet-, and adipose-like cells.
METHODS: Human umbilical cord venous endothelial and subendothelial layer cells were isolated and grouped according to different implantation densities for primary culture. The time for primary culture was recorded and cell passage was performed. Passage 1 human umbilical cord mesenchymal stem cells were in vitro induced to differentiate into neuron-, islet-, and adipose-like cells.
RESULTS AND CONCLUSION: Human umbilical cord mensenchymal stem cells at an implantation density of 5×105-1 ×106/cm2 cell density showed optimal growth. After induced by β-sulfhydryl alcohol for 6 hours, human umbilical cord mesenchymal stem cells expressed nestin, neurofilament protein and glial fibrillary acidic protein. After induced by niacinamide, activin A, glucagon-like peptide 1 for 21 days, obvious islet-like cell masses appeared and were stained red by dithizone. After induced by adipogenic culture medium, cells were positive for oil red O staining, with appearance of obvious liquid drops. A large number of human umbilical cord mesenchymal stem cells could be quickly harvested at a proper implantation density and the cells could be induced to differentiate into neuron-, islet-, and adipose-like cells.