Microcarrier cultivation of silkworm BmN cell and high-level expression of HBeAg gene

DENG Xiao-Zhao; DIAO Zhen-Yu; ZHU Ying; HE Liang; JIAO Ren-Liang; ZHOU Zong-An; ZHANG Lin-Yuan

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June 1998

DENG Xiao-Zhao, DIAO Zhen-Yu, ZHU Ying, HE Liang, JIAO Ren-Liang, ZHOU Zong-An and ZHANG Lin-Yuan. Microcarrier cultivation of silkworm BmN cell and high-level expression of HBeAg gene[J]. Virologica Sinica, 1998, 13(3).

Citation: DENG Xiao-Zhao, DIAO Zhen-Yu, ZHU Ying, HE Liang, JIAO Ren-Liang, ZHOU Zong-An, ZHANG Lin-Yuan. Microcarrier cultivation of silkworm BmN cell and high-level expression of HBeAg gene .VIROLOGICA SINICA, 1998, 13(3) : 237.

家蚕BmN细胞的微载体培养及HBeAg的高效表达

1.
南京军区后勤部军事医学研究所分子生物学研究室

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３．６１０５细胞／ｍＬ、微载体浓度５ｇ／Ｌ），５ｄ后，细胞的终密度达到２．８１０６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∶９．６１０４，是用方瓶静止培养的家蚕细胞表达量的３倍。
- 微载体昆虫细胞培养乙肝病毒E抗原基因表达

Microcarrier cultivation of silkworm BmN cell and high-level expression of HBeAg gene

Abstract

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 1.010 5 cells/mL when Cytodex 3 microcarrier concentration was 3g/L. Cultured under the proper conditions (inoculation concentration: 3.610 5 cells/mL; microcarrier cytodex 3 concentration: 5g/L) for 5 days, the final cell concentration of BmN was 2.810 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.
- Microcarrier Insect cell culture HBeAg Gene expression

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Microcarrier cultivation of silkworm BmN cell and high-level expression of HBeAg gene

Abstract: 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 1.010 5 cells/mL when Cytodex 3 microcarrier concentration was 3g/L. Cultured under the proper conditions (inoculation concentration: 3.610 5 cells/mL; microcarrier cytodex 3 concentration: 5g/L) for 5 days, the final cell concentration of BmN was 2.810 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.