Observation of human parotid gland epithelial cells of continuous passage culture in vitro
- VernacularTitle:体外连续传代培养人腮腺腺上皮细胞的观察
- Author:
Linpu ZHANG
;
Guanhua WANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
The experiment was conducted in the Central Laboratory of Tianjin Stomatological Hospital from November 2004 to March 2005. Epithelial cells were isolated from normal parotid gland tissues obtained form resected benign tumor of an adult, so as to prepare rat tail collagen. Two adult rats were selected to obtain the tendon fascia from rat tail, which were then immersed in the 500 mL of 0.1% glacial acetic acid. The infiltration culture board of collagen glacial acetic acid, proximal wall of culture flask and beaker with ammonia water were placed in a sterile containers to reserve at 37 ℃ for 72 hours. The epithelial cells were isolated from parotid gland tissues by enzyme digestion and cultured in 1:1 DMEM/F12 culture medium supplemented with some growth stimulating factors such as insulin (INS), hydrocortisone (HC) and isoproterenol (ISO) by using self-made rat tail collagen gel substrate. The cytomorphological characteristics of primary and passage cells were observed with inverted microscope. The result showed that the primary culture of parotid gland epithelial cells: cells were in polarity arrangement on the 4th day and formed in different size of acinus and pip kind structure. The serial subcultivation of parotid gland epithelial cells. In the culture period of 50 days, parotid gland epithelial cells were passed to the F3 generation, and the cells of F3 generation frozen in liquid nitrogen recovered and survived. It could be seen by HE staining that the cell body was bigger, the kytoplasm was abundant and the nuclear membrane was clear with one or two entoblasts. The karyogenetic division could be found in partial entoblast, whereas no abnormal karyogenetic division was seen.
