[Cellular phenotype conversion induced by co-culture of human mesenchymal stem cells cocultured with human sweat gland cells].
OBJECTIVE
To investigate the cellular phenotype conversion during human mesenchymal stem cells (hMSCs) cocultured with injured human sweat gland cells (hSGCs) in vitro.


METHODS
HMSCs and hSGCs were isolated and cultured and expanded respectively. The antigens expression of hMSCs and hSGCs were detected by two-steps immunocytochemistry. HMSCs were labeled with BrdU. The hSGCs were heat-shocked at 47 degrees C for 40 min when they reached 70% confluency, then cooled for 1-2 h at 37 degrees C and (1 - 2) x 10(5) BrdU-labeled hMSCs were added before incubation for up to 2 weeks. The cocultures were observed by phase contrast microscopy and detected by double-staining immunocytochemistry using CEA and BrdU as primary antibodies.


RESULTS
The cultured hMSCs and hSGCs were clonogenic growth. HMSCs were positive for anti-CD44 and anti-CD105 staining and negative for anti-CD34 and anti-CEA staining. HSGCs express CK7, CK18, CK19 and CEA. The positive rate of BrdU labeled-hMSCs was 90%. The majority of hSGCs lost cell-cell contact after heat-shock. 2 weeks after cocultured, some cocultured cells were positive for both anti-CEA and anti-BrdU staining and some cocultures had more than two nuclei which stained with two different colors by double-staining immunocytochemistry. Statistic results showed 1%-5% of the hMSCs added to the coculture system were recovered as double-staining cells expressing BrdU and CEA while only 0.01%-0.05% cells stained with two different colors in nuclei. The multi-nucleated cells were wide and flatten.


CONCLUSIONS
HMSCs could differentiate into hSGCs in vitro under injured microenvironment. The mechanisms of which may be that hMSCs differentiate into hSGCs directly or by cell fusion, even nucleus fusion.
Li
Hai-hong
HH
Key Research Laboratory of the Wound Repair, the 304th Clinical Department of General Hospital of PLA, Beijing 100037, China.
Fu
Xiao-bing
XB
Zhou
Gang
G
Fei
Pei
P
Chen
Wei
W
Bai
Xiao-dong
XD
Cai
Cun-liang
CL
Sun
Tong-zhu
TZ
D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D002459:Cell Fusion; D002478:Cells, Cultured; D018920:Coculture Techniques; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010641:Phenotype; D013545:Sweat Glands / Q000166:cytology*