摘要
选择微载体Ｃｙｔｏｄｅｘ３对家蚕ＢｍＮ（从ＳｉｌｋｗｏｒｍＢｏｍｂｙｘｍｏｒｉ获得的细胞系）细胞进行高密度培养，并获得成功。观察了家蚕ＢｍＮ细胞在微载体Ｃｙｔｏｄｅｘ３上的贴壁分布，研究了不同接种浓度与微载体浓度时细胞的生长情况，并筛选出最佳的技术参数。在１０００ｍＬ滚瓶中用微载体Ｇｙｔｏｄｅｘ３培养家蚕细胞，在合适的培养条件下（细胞接种浓度３．６×１０５细胞／ｍＬ、微载体浓度５ｇ／Ｌ），５ｄ后，细胞的终密度达到２．８×１０６细胞／ｍＬ，细胞增长指数为７．９。在细胞指数生长期（传代后４８～６０ｈ）用载有ＨＢｅＡｇ基因的重组病毒（ｒＢｍＨＢｅ）接种（感染量为０．４ＰＦＵ／细胞），５ｄ后，培养上清中ＨＢｅＡｇ滴度为１∶９．６×１０４，是用方瓶静止培养的家蚕细胞表达量的３倍。
Distribution of silkworm Bm N cell attachment to Cytodex 3 microcarrier was observed and the influence of cell inoculation concentration and microcarrier concentration on cell growth was studied. The minimal cell inoculation concentration was 1.0×10 5 cells/mL when Cytodex 3 microcarrier concentration was 3g/L. Cultured under the proper conditions (inoculation concentration: 3.6×10 5 cells/mL; microcarrier cytodex 3 concentration: 5g/L) for 5 days, the final cell concentration of BmN was 2.8×10 6 cells/mL. The cell growth index was 7.9. At 48-60 hours after cell passage, the Bm N cells was inoculated with rBmHBe (multiplicity of infection Mol=0.4 PFU/cell). Five days later the HBeAg titer in the medium was 1∶96000 and in the control group it was 1∶32000.
出处
《中国病毒学》
CSCD
1998年第3期237-242,共6页
Virologica Sinica
基金
江苏省自然科学基金
