Study on microencapsulated human chromaffin cells cultured in vitro / 中华麻醉学杂志

ABSTRACT

Objective To compare the catecholamine (CA ) and M enkephalin(M ENK) release from human chromaffin cells (HCC) and microencapsulated HCC (ME HCC ) and investigate the effects of microencapsulation on the growth and activity of chromaffin cells Methods Adrenal glands were taken from brain death healthy adults The chromaffin cells were isolated and primarily cultured in vitro During culture the chromaffin cells were HE stained and assessed by labelling the cells with tyrosine hydroxylase monoclonal antibody The percentage of tyrosine positive cells were counted under microscope (1) The chromaffin cells were microencapsulated with 2% alginate and cultured in vitro (experimental group) HCC which were not microencapsulated were used as control The culture media of both groups were replaced every 48h and collected and stored under -20℃ for determination of CA and M ENK concentration (2) On the 6th day of culture, nicotine was added to HCC and MC HCC suspension After 30 min incubation, the suspension was centrifuged and the supernate collected and stored under -20 ℃ for determinatoin of CA and M ENK concentration with radioimmunoassay Results (1) ME HCC grew fairly well in vitro in the culture medium and was morphologically similar to HCC (2) There was no significant difference in CA and M ENK concentration in HCC and ME HCC culture media (3) CA and M ENK concentrations in supernate were increased by nicotine stimulation and there was no difference in the CA and M ENK concentration in the supernate between the two groups Conclusions Alginate and microencapsulation technique are not harmful to HCC, ME HCC has fairly good activity and release function and can be effectively used for transplantation