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PK15细胞的无血清全悬浮驯化研究
刘天伦,郎洪彬,孔飒
0
(北京民海生物科技有限公司)
摘要:
目前PK15细胞主要用于生产猪圆环病毒2型,传统的培养工艺为滚瓶系统或微载体系统,该两种培养系统都存在各自的劣势。驯化一株全悬浮的PK15细胞可推进工艺的进一步升级,降低成本的同时也简化了培养工艺且更易于产业化。经贴壁降血清培养、低血清悬浮优化传代、无血清悬浮传代培养, 对一株贴壁的PK15细胞进行了无血清的全悬浮驯化。传代23次后,该株细胞无血清全悬浮培养初始密度为0.5×106/mL,在72 h密度可增殖到4.0×106/mL左右,形态良好,倍增时间稳定,且细胞活率达到95%以上,命名为PK15-S,为大规模培养PK15细胞提供了理论依据。
关键词:  PK15细胞  细胞驯化  无血清  全悬浮  
DOI:
投稿时间:2019-07-07修订日期:2019-08-28
基金项目:
Domestication of a PK15 cell line to suspension culture by serum-free medium
(Beijing Minhai Biotechnology Co., Ltd.)
Abstract:
Abstract: Currently, PK15 cells are?mainly used to culture porcine circovirus type 2. The traditional method for the cultivation of PCV-2 was using the rolling bottle system or micro carrier system. However, both of those systems have their own disadvantages. Hence, domesticate a?suspension PK15 cell line can promote the upgrading of the traditional methods, and also can?reduce costs. Meanwhile, it can also simplify the culture process to be?easier to industrialization. In this study, an adherent PK15 cell line was domesticated as a suspension cell line in serum-free medium. After 23 passages, the density of the domesticated cell line could increase from the initial density (0.5×106/mL) to 4.0×106/mL in 72 h. Furthermore, the cell morphology is good, the cell proliferation time is stable, and the cell survival rate is above 95%. In conclusion, this study provides a theoretical basis for large-scale culture of PK15 cells.
Key words:  PK15 cells  domestication  serum free  suspension

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