Dissociation, culture and morphologic changes of interstitial cells of Cajal in vitro.
OBJECTIVE
To study the method of dissociation, culture and investigate its morphologic changes in vitro of interstitial cells of Cajal (ICC).


METHODS
Enzymatic digestion and Ficoll density centrifugation were used to dissociate ICC from the ileal segment of mice. Factors including contamination, Ca(2+), Mg(2+) and collagenase, and stem cell factor, etc., were investigated. ACK2, the antibody of c-kit, was used to identify the cultured ICC. Both light microscope and fluorescence microscope were used to observe the changes of ICC in vitro.


RESULTS
The method for dissociation and culture of ICC in vitro was successfully established. After 24 h, cultured ICC exhibited a few axis-cylinders, and longer axis-cylinders were observed to form synapse of each other after 3 d. More widespread connections formed within 7 d in vitro. The changes of its morphologic character were obvious within 7 d; however, there were no obvious morphologic changes after 30 d.


CONCLUSIONS
Many factors can influence the dissociation and culture of ICC.
Li
Chun-Xue
CX
Department of General Surgery, Daping Hospital, Third Military Medical University, Chongqing 400042, China. lichunxue1974@hotmail.com
Liu
Bao-Hua
BH
Tong
Wei-Dong
WD
Zhang
Lian-Yang
LY
Jiang
Yan-Ping
YP
D000818:Animals; D001683:Biological Clocks; D018929:Cell Culture Techniques; D002469:Cell Separation; D002478:Cells, Cultured; D041981:Gastrointestinal Tract / Q000166:cytology* / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009130:Muscle, Smooth / Q000166:cytology* / Q000502:physiology*; D013997:Time Factors