Culture of tissue-engineered human normal esophageal epithelial cells
- VernacularTitle:组织工程化人正常食管上皮细胞的培养
- Author:
Qizhou BAI
;
Yongzhu YANG
;
Zhiqiang WANG
;
Yunjiu GOU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(33):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: In vitro isolation and culture of esophageal epithelial cells are basic component of tissue-engineered esophagus. This study explored the method to culture esophageal epithelial cells for research on tissue-engineered esophagus. METHODS: The experiment was performed at experimental center of Lanzhou University from May to November 2007. Normal esophagus tissues, 2.0-3.0 cm, were harvested from patient with esophagus cancer by surgery. The informed consent was obtained from the patient. Esophageal epithelial cells for tissue engineering were obtained and passaged. The cells cultured by DMEM+F12 (1∶1) after 20 minutes, 1-4 days were observed by immunohistochemistry staining and inverted phase contrast microscope. The growth curve of cells was drawn by MTT method. RESULTS: The immunohistochemistry staining results showed that 90% of the cells were positive, which indicated the cultured cells were esophageal epithelial cells. Normal cells were big and globular, floating in the culture-medium. Cells began to adhere after 20 minutes, and most cells were polygon-like or irregular globular and adherent after 1 day; the cells began to cluster after 2 days; the cells grew at peak after about 3-4 days with abundant endochylema and large and spherical nuclear. Cells growth reached the peak after about 3 days of culture and its absorbance was significantly different compared with that on the 1st, 2nd, 5th, and 6th days (P