The Improvement in the Primary Culture of Alveolar Epithelial Type of Rats

【Abstract】 Objective To improve the method of culturing and obtain purified alveolar epithelial typeⅠ (ATⅠ) cells of SD rats. Methods 1 d newborn SD rats were applied for cell culture and brains were decapitated for lung tissues obtaining after respiratory cessation. CollagenaseⅠand DNaseⅠ were used to digest and isolate cells. Then, cells were put into the plate coated with typeⅠ rat tail collagen and different adherence was used to purify cells. Meanwhile, culture medium replacement was conducted after 48 h. The growth status was observed under an inverted microscope. Immunofluorescence including specific marker AQP5, SPC, BSI, Vimentin were used to identify cells. Results 2 d after incubation, the cells began to adhere to plate. At day 4 and 6, cells began to proliferate and exhibited a shape of spindle, cube and polygon. 8 d after incubation, the character of proliferation reached the highest and the cell viability was (87±8)% and purification was (87±5)%. Conclusion By modifying the methods of tissues harvest, isolation and culture, we can obtain AT Ⅰ cells with high yield, good growth state and super purity.