Evaluation of royal jelly as an alternative to fetal bovine serum in cell culture using cell proliferation assays and live cell imaging

Abstract

Background: Royal jelly is a nutritious substance produced by the young nurse bees and contains significant amounts of proteins which are important for cell growth and proliferation. The aim of this study was to evaluate the effect of royal jelly as an alternative to fetal bovine serum (FBS) in cell culture using cell proliferation assays and live cell imaging. Materials and Methods: MRC-5 cells were treated with various concentrations of royal jelly extract in MTT assay. The control groups were comprised of Alpha-Minimal Essential Medium (α-MEM) alone and α-MEM with 10% FBS. Subsequently, the cell proliferation was studied for 10 days using Alamar Blue assay and live cell imaging from 48 to 72 h. The population doubling time (PDT) was determined using trypan blue assay after live cell imaging. Results: In MTT assay, 0.156 and 0.078 mg/ml of royal jelly produced higher cell viability compared to positive control group but were not significantly different (P > 0.05). In the Alamar Blue assay, 0.156 and 0.078 mg/ml of royal jelly produced greater percentage of reduction at day 3 even though no significant difference was found (P > 0.05). Based on live cell imaging, the PDT for positive, negative, 0.156 and 0.078 mg/ml of royal jelly groups were 29.09, 62.50, 41.67 and 41.67 h respectively. No significant difference was found in the PDT between all the groups (P > 0.05). Conclusion: Royal jelly does not exhibit similar ability like FBS to facilitate cell growth under the present test conditions.

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

“…A lower temperature is essential to maintain the activity of MRJPs. The exposure of RJ extract to 25 kGy γ radiation may be the main cause of RJ loss of cell growth activity in the report of Musa et al (2013). In our investigation, the protocols of the extract, purification, and qualification of MRJPs have been standardized, which could ensure objective results in culture cells.…”

“…The evaluation of RJ extract or RJ proteins was implemented in several cell lines from insects, rats, and humans (Kimura Y. et al, 1996;Kimura M. et al, 2003;Salazar-Olivo and PazGonzalez, 2005). However, most previous work (Musa et al, 2013;Yu et al, 2015) focused on using single RJ extract or RJ proteins to replace FBS. The cell type and the combination formula of RJ extract or RJ proteins with FBS and cytokines have been neglected (Musa et al, 2013).…”

“…However, most previous work (Musa et al, 2013;Yu et al, 2015) focused on using single RJ extract or RJ proteins to replace FBS. The cell type and the combination formula of RJ extract or RJ proteins with FBS and cytokines have been neglected (Musa et al, 2013). In addition, more evidence has shown that MRJPs, especially MRJP1, are the key active factors for honeybee cast variation and larvae development, and animal and human cells.…”

“…For example, a recent investigation showed that RJ extract does not exhibit the activity in MRC-5 cells, human fibroblast cells (Musa et al, 2013). In addition, differential effects of MRJPs in combination with different cytokines on cell viability remain unclear.…”

“…For this reason, development of alternatives to replace FBS is essential, particularly for cell culture purposes. Hence, more investigations to evaluate the activities of RJ or RJ proteins have been carried out (Karaca et al, 2012;Musa et al, 2013). Kimura et al (2003) found that 350-kDa RJ glycoprotein stimulates proliferation of human monocytes.…”