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Journal of Zhejiang University SCIENCE B

ISSN 1673-1581(Print), 1862-1783(Online), Monthly

Evaluation of the major royal jelly proteins as an alternative to fetal bovine serum in culturing human cell lines

Abstract: Royal jelly (RJ) is a well-known bioactive substance. It contains large amounts of major royal jelly proteins (MRJPs), which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum (FBS) in different types of human cell culture, the proliferation of the complex serum with different ratios of MRJPs/FBS (M/F) was evaluated on five cell lines: 293T, HFL-I, 231, HCT116, and Changliver using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor (EGF) and insulin-transferrin-selenium (ITS), promoted proliferations of Changliver, 293T, HCT116, and HFL-I by 18.73%‒56.19% (P<0.01). Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.

Key words: Major royal jelly proteins (MRJPs), Cell culture, Alternative, Fetal bovine serum (FBS), Cytokine

Chinese Summary  <30> 蜂王浆主蛋白(MRJPs)作为灭活胎牛血清(FBS)替代品培养人体细胞的效果评价

目的:为MRJPs应用于人体细胞培养提供科学依据,为发展蜂王浆在细胞工程的新用途提供技术支撑。
创新点:首次验证MRJPs对多种人体细胞的促分裂效果,证实MRJPs可部分替代FBS培养人体细胞,并取得培养人体细胞的MRJPs与FBS和细胞因子的适合比例。
方法:鲜蜂王浆中分离得到的MRJPs溶液与FBS按不同比例(0/10、3/7、6/4和9/1)复配成复合血清,分别添加于无血清培养基中用于培养293T、HFL-I、231、HCT116和Changliver等5种人体细胞。以添加体积分数为10%的FBS的无血清培养基为对照,根据MTT法测定的吸光度和细胞形态比较分析结果,得到促进细胞分裂的MRJPs溶液与FBS最佳配合比例。然后在筛选出的最佳复合血清中添加不同的细胞因子,再通过同样的比较分析,得到适合的细胞因子组合。
结论:MRJPs对多种人体细胞具有促进分裂作用,可部分替代FBS培养人体细胞。MRJPs与FBS的最佳配比为:60% MRJPs溶液和40% FBS(复合血清);该复合血清与细胞因子的最佳组合为:复合血清+表皮生长因子+胰岛素-转铁蛋白-硒。

关键词组:蜂王浆主蛋白;灭活胎牛血清;细胞培养基;替代品;分裂; 细胞因子


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DOI:

10.1631/jzus.B1500295

CLC number:

Q253; S896.3

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On-line Access:

2024-08-27

Received:

2023-10-17

Revision Accepted:

2024-05-08

Crosschecked:

2016-05-12

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