1.Study on the differentiation of human umbilical cord perivascular cell into cardiac-like cells in vitro.
Yu HU ; Tie-Bi TONG ; Li DU ; Al ET ;
Chinese Journal of Practical Internal Medicine 2006;0(S1):-
Objective To investigate the capability of cardiomyoganic differentiation of human umbilical cord perivascu- lar cells induced by 5-azacitidine in vitro.Methods Human umbilical cord perivascular colls were harvested from the umbilical cords of consenting full-term caesarian section patients,which were digested by collagenase,and were purified, expanded through repeating passaging in time in vitro.The HUCPV cell of passage 3 were treated with 5-azacytidine(5 -aza,5?10~(-6)nmol/L) for 24 hours and continued to be cultured under the previous condition for 4 weeks.The prolifera- tion and differentiated phenotype of HUCPV cells were observed.Results The isolation method of the UC vasculature and enzymatic digestion of its perivascular tissue to rapidly harvest a highly proliferative HUCPV cells.After treated with 5 -aza for 4 weeks,the passage 3 HUCPV cells formed the spindle-shaped cells and rate of nuclear and cytoplasm de- creased.Myotube-like structures were found in some cytoplasm.Immunohistochemistry results showed that the HUCPV cells were similar to MSC by stained positively for CD44 and negtively for CD34.The HUCPV cells induced by 5-aza were stained positively a-cardiac action and cardiac Troponin by immunohistochemistry.Furthermore,the expression of a -cardiac action and cardiac treponin gene as detected positively by RT-PCR after HUCPV cells were induced by 5- aza.Conclusion The HUCPV cell was a valuable material for cell-based therapies and can be induced into cardiac- like cells by treatment with 5-aza.