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. 2016 Jun;17(6):476-83.
doi: 10.1631/jzus.B1500295.

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

Di Chen  1 , Xiao-Xuan Xin  1 , Hao-Cheng Qian  1 , Zhang-Yin Yu  1 , Li-Rong Shen  1
Affiliations

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

Di Chen et al. J Zhejiang Univ Sci B. 2016 Jun.

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.

Keywords: Alternative; Cell culture; Cytokine; Fetal bovine serum (FBS); Major royal jelly proteins (MRJPs).

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Conflict of interest statement

Compliance with ethics guidelines: Di CHEN, Xiao-xuan XIN, Hao-cheng QIAN, Zhang-yin YU, and Li-rong SHEN declare that they have no conflict of interest.

This article does not contain any studies with human or animal subjects performed by any of the authors.

Figures

Fig. 1
Fig. 1
SDS-PAGE analysis of MRJPs used in culturing test cell lines Lanes of M and 1 refer to protein marker and MRJPs, respectively
Fig. 2
Fig. 2
Cellular morphology of 293T and HCT116 (on Day 5) treated by the complex M/F 6/4 serum and FBS, respectively (a) 293T+FBS; (b) 293T+M/F 6/4; (c) HCT116+FBS; (d) HCT116+M/F 6/4
Fig. 3
Fig. 3
Effects of several cytokines together with the complex M/F 6/4 serum on the proliferations of 231 (a) and Changliver (b) cells (on Day 3) The means of absorbance denoted with the same letter (a‒g) do not differ significantly from each other. Data are expressed as mean±SD (n=3, 4)
Fig. 4
Fig. 4
Cellular morphology and densities of Changliver (on Day 3) cultured with complex M/F 6/4 serum (FM) and the combination of FM and cytokines ITS+EGF (FMIE), respectively (a) Changliver+FM; (b) Changliver+FMIE
Fig. 5
Fig. 5
Effects of combinations of cytokines together with the complex M/F 6/4 serum on the proliferation of 293T cells (a) (on Day 3) and HCT116 cells (b) (on Day 5) The means of absorbance denoted with the same letter (a‒c) do not differ significantly from each other. Data are expressed as mean±SD (n=3, 4)
Fig. 6
Fig. 6
Effects of combinations of cytokines together with the complex M/F 6/4 serum on the proliferation of HFL-I cells (on Day 10) The means of absorbance denoted with the same letter (a‒c) do not differ significantly from each other. Data are expressed as mean±SD (n=3, 4)
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References

    1. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72(1-2):248–254. (Available from: http://dx.doi.org/10.1016/0003-2697(76)90527-3) - DOI - PubMed
    1. Buttstedt A, Moritz RF, Erler S. Origin and function of the major royal jelly proteins of the honeybee (Apis mellifera) as members of the yellow gene family. Biol Rev. 2014;89(2):255–269. (Available from: http://dx.doi.org/10.1111/brv.12052) - DOI - PubMed
    1. Chen G, Yue A, Ruan Z, et al. Human umbilical cord-derived mesenchymal stem cells do not undergo malignant transformation during long-term culturing in serum-free medium. PLoS ONE. 2014;9(6):e98565. (Available from: http://dx.doi.org/10.1371/journal.pone.0098565) - DOI - PMC - PubMed
    1. Drapeau MD, Albert S, Kucharski R, et al. Evolution of the Yellow/Major Royal Jelly Protein family and the emergence of social behavior in honey bees. Genome Res. 2006;16(11):1385–1394. (Available from: http://dx.doi.org/10.1101/gr.5012006) - DOI - PMC - PubMed
    1. EL-Ensahsy HA, Abdeen A, Abdeen S, et al. Serum concentration effects on the kinetics and metabolism of HeLa-S3 cell growth and cell adaptability for successful proliferation in serum free medium. World Appl Sci J. 2009;6(5):608–615.