关键词: 细胞培养, 细胞增殖

Abstract: Abstract:Objective To establish the culture methods of submandibular gland cells from Wistar rats in vitro and lay a foundation for amplification of submandibular gland cells and gene therapy of salivary diseases.Methods The submandibular gland was obtained from Wistar rats which were one day old,after digested with LB3 and pancreatin，the submandibular gland cells were cultivated in DMEM/F12 culture fluid containing 2% fetal calf serum,epidermal gowth factor and insulin.The morphology of the culture and subculture cells was observed by phase contract microscope.The cell origin was identified by immunohistostaining.PAS staining was used to observe cell function of synthesis and secretion of polysaccharides.Results The morphous of primary cultured cells was round,triangle or polygon.At passage 2,the cells grew well,there was no change of morphous.The cells were positively stained with cytokeratin,E- cadherin and PAS,it indicated that the cells were epithelium origin and had the function of synthesis and secretion of polysaccharides.Conclusion The Wistar rat submandibular gland cells are successfully isolated and cultivated with enzyme digestion.The cultured submandibular gland cells at passage 2 still maintain their biocharacteristics.

Key words: cell culture, cell proliferation