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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2018, Vol. 44 Issue (1): 1-9    DOI: 10.3785/j.issn.1008-9209.2017.06.281
生物科学与技术     
王浆主蛋白作为胎牛血清替代物培养中国仓鼠卵巢细胞CHO-K1的效果及作用机制
钱浩诚1，蒋晨旻1，陈华才2，沈立荣1*
1. 浙江大学生物系统工程与食品科学学院，馥莉食品科学研究院，浙江省农产品加工技术研究重点实验室，杭州 310058；2.中国计量大学光学与电子科技学院，杭州 310018
Effect and mechanism of major royal jelly proteins as an alternative of fetal bovine serum to culture Chinese hamster ovary cell CHO-K1
QIAN Haocheng1, JIANG Chenmin1, CHEN Huacai2, SHEN Lirong1*
1. Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China; 2. College of Optical and Electronic Technology, China Jiliang University, Hangzhou 310018, China
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摘要：

用王浆主蛋白（major royal jelly proteins, MRJPs）部分代替胎牛血清（fetal bovine serum, FBS）培养中国仓鼠卵巢细胞（Chinese hamster ovary cell, CHO-K1），比较添加不同MRJPs/FBS（M/F）比例的培养基对细胞相对增殖率的影响。四唑盐比色法（methyl thiazolyl tetrazolium, MTT）结果显示：当M/F 比例为50/50 时对细胞的促增殖作用最显著；与完全培养基对照组（M/F 0/100）相比，培养48 与72 h 时的相对活细胞数分别上升了25.1%和13.6%。细胞成像仪分析表明，M/F 50/50 培养基的细胞密度最高，其细胞直径显著大于完全培养基对照组。流式细胞仪结果显示，与完全培养基对照组相比，M/F 50/50 培养基中的细胞增殖指数（proliferation index, PI）提高了19.4%，推测MRJPs的促细胞增殖作用可能与DNA、蛋白质及酶的合成有关。拉曼光谱检测结果显示，各处理组培养基的细胞光谱图之间没有显著差异，说明MRJPs在促进细胞增殖的同时维持了细胞的稳定性。以上结果证明，MRJPs可以部分替代FBS培养生物医药产业中应用广泛的CHO-K1细胞，降低细胞培养成本，具有产业化前景，并为蜂王浆深加工提供新的用途。
关键词： 王浆主蛋白;  中国仓鼠卵巢细胞;  促增殖;  细胞周期;  胎牛血清;  拉曼光谱    
Abstract:

The major royal jelly proteins (MRJPs) as the alternative of fetal bovine serum (FBS) were used to culture Chinese hamster ovary cell line (CHO-K1). The effects of mixture media with different MRJPs/FBS (M/F) proportions on the proliferation of CHO-K1 cell were compared. The result of methyl thiazolyl tetrazolium (MTT) assay showed that the cells in the medium with M/F 50/50 possessed the highest relative proliferation activity. Compared to the CK (M/F 0/100), the relative cell proliferation activity in the medium with M/F 50/50 was increased by 25.1% (48 h) and 13.6% (72 h), respectively. Cell imager showed that the cell density and diameter in the medium with M/F 50/50 were significantly higher than those in the CK. The detection of cell cycle with flow cytometry showed that the cell proliferation index (PI) in the medium with M/F 50/50 was increased by 19.4% compared to that in the CK, indicating that MRJPs might promote the synthesis of DNA, proteins and enzymes. The Raman spectrum of cells in the medium with M/F 50/50 was similar with those in the CK and bovine serum albumin (BSA) medium, demonstrating that the complex serum maintains the cell type stable. In conclusion, the results suggest that MRJPs could partially replace FBS in the cultivation of CHO-K1 cell line and possess industry prospect.
Key words: Chinese hamster ovary cell    fetal bovine serum    Raman spectrum    cell cycle    major royal jelly protein    proliferation
收稿日期: 2017-06-28 出版日期: 2017-09-06
CLC:  Q 253  
基金资助: 国家自然科学基金面上项目（31271848）；浙江省科技计划项目（2017C32033）；杭州市重大创新科技项目（20131812A25)
通讯作者: 沈立荣（http://orcid.org/0000-0001-8240-1286）     E-mail: shenlirong@zju.edu.cn
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引用本文:

钱浩诚，蒋晨旻，陈华才，沈立荣. 王浆主蛋白作为胎牛血清替代物培养中国仓鼠卵巢细胞CHO-K1的效果及作用机制[J]. 浙江大学学报(农业与生命科学版), 2018, 44(1): 1-9.

QIAN Haocheng, JIANG Chenmin, CHEN Huacai, SHEN Lirong. Effect and mechanism of major royal jelly proteins as an alternative of fetal bovine serum to culture Chinese hamster ovary cell CHO-K1. Journal of Zhejiang University (Agriculture and Life Sciences), 2018, 44(1): 1-9.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2017.06.281    或    http://www.zjujournals.com/agr/CN/Y2018/V44/I1/1

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