[Experimental study on biological character changes of human epidermal cells during proliferation culture in vitro].
[Abstract]


OBJECTIVE
The study was (1) to investigate the biological character changes of human epidermal cells during proliferation culture in vitro and (2) to provide data for construction of engineered skin.


METHODS
The foreskin was collected from 20 healthy children. The epidermal cells were isolated with digestion of the foreskin and cultured in vitro. Growth curve was obtained from the data of cell counting. Cell growth kinetics was observed. Meanwhile, clonal analysis and cell size measurement was performed. The rate of keratin 19 (K19) and involucrin expression-positive cells was counted by flow cytometer. Expression of K19 and involucrin mRNA was detected by RT-PCR.


RESULTS
When passaged to P6, the keratinocytes from child foreskin could be expanded by (700 +/- 37) times. Flow cytometer results showed that the percentage of K19 expression-positive cells decreased from (66.97 +/- 3.14)% to (4.65 +/- 1.38)% while the percentage of involucrin expression-positive cells increased from (11.65 +/- 1.62)% to (97.03 +/- 2.66)% at P0 and P6 respectively. RT-PCR results showed that expression of K19 mRNA decreased from P0 to P6 while involucrin mRNA kept stable with passage in vitro.


CONCLUSIONS
Human epidermal cells of passage 5 maintain proliferation phenotype, which are suitable for skin tissue engineering. Decrease of proliferation phenotype content is partially responsible for the proliferation capacity loss of in vitro cultured epidermal cells.
Deng
Chen-liang
CL
Department of Plastic Surgery, Shanghai Ninth People's Hospital, Shanghai Tissue Engineering Research & Jevelopment Center, Shanghai 200011, China.
Cui
Lei
L
Yang
Guang-hui
GH
Yang
Jun
J
Liu
Wei
W
Cao
Yi-lin
YL
D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D000078404:Epidermal Cells; D052816:Foreskin / Q000166:cytology; D006801:Humans; D008297:Male; D023822:Tissue Engineering / Q000379:methods*