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.

Anti-senescence effect and molecular mechanism of the major royal jelly proteins on human embryonic lung fibroblast (HFL-I) cell line

“…HFL-I is also used as a model organism in cell apoptosis tests (Xu et al, 2012;Ren et al, 2013;Chen et al, 2014;Ershova et al, 2016) and proliferation tests (Vietti et al, 2013;Zeng et al, 2015). We previously found that FBS culture media partially replaced by MRJPs could exert a proliferation effect on several cell lines, including the HFL-I cell line (Chen et al, 2016). Cell senescence is followed by a series of phenomena, including morphological changes, the loss of proliferation ability, and an increase in SA-β-gal activity (Cristofalo and Pignolo, 1993;Dimri et al, 1995;Toussaint et al, 2000;Liu et al, 2007;Debacq-Chainiaux et al, 2008;Hwang et al, 2009).…”

“…MRJPs extracted from RJ can stimulate the proliferation of several cell lines, including hepatocytes (Kamakura et al, 2001), human myeloid cell line U-937 (Watanabe et al, 1998), human monocytes (Kimura et al, 2003), and Tn-5B1-4 insect cells (Salazar-Olivo and Paz-González, 2005). Our recent study suggests that MRJPs can increase mean lifespan and reproductivity in Drosophila (Xin et al, 2016) and partially replace fetal bovine serum (FBS) for culturing several human cell lines and increase the cell viability (Chen et al, 2016).…”

Multifunctionality and intrinsic disorder of royal jelly proteome

“…[80], whereas Chen et al. [199] reported that MRJPs are involved in the increase of cell viability, and can be used for partial replacement of the fetal bovine serum (FBS) during culturing of several human cell lines. MRJPs demonstrated the anti‐senescence effect on human embryonic lung fibroblast (HFL‐I) cell line, likely via the upregulation of the expression of superoxide dismutase‐1 (SOD1) and downregulation of the expression of mammalian target of catenin beta like‐1 (CTNNB1), rapamycin (MTOR), and tumor protein p53 (TP53) [200].…”

Partial recovery of MRJP1 protein expressed in Pichia pastoris using chromatographic techniques

“…Major Royal Jelly Protein 1 (MRJP1), also called royalactin, is the major protein component of the beeproduced complex mixture royal jelly (Shen et al, 2015;Ibarra-Herrera et al, 2014, Buttstedt et al, 2014. It has been reported that MRJP1 promotes health benefits spanning various species including the mouse Mus musculus, the rat Rattus norvegicus, the fruit fly Drosophila melanogaster and the worm Caenorhabditis elegans (Kamakura b et al, 2001;Detienne et al, 2014;Shorter et al, 2015;Chen et al, 2016;Xin et al, 2016).…”