5.1 Cell source and culture conditions of RBL-2H3 cell line
Most of the RBL cell lines used in allergy research were the RBL-2H3 cell line (131 of 134), followed by humanized RBL cells (3 of 134). The American Type Culture Collection (ATCC) is the most reported source (46 of 131) for the RBL-2H3 cell line. Other cell bank sources of RBL-2H3 includes China Center for Type Culture Collection (CCTCC) (20 of 131), Japanese Collection of Research Bioresources Cell Bank (JCRB) (9 of 131), Korean Cell Line Bank (KCLB) (7 of 131), Procell Life Science & Technology (PLCT) (3 of 131), Bioresource Collection and Research Center (BCRC) (2 of 131), Chinese Academy of Sciences Shanghai (CAS) (2 of 131), Binsui Biotechnology (SBB) (1 of 131), Cellcook Biotechnology (CB) (1 of 131), Shanghai EK-Bioscience Biotechnology (SEB) (1 of 131), National Infrastructure of Cell Line Resource (NICLR) (1 of 131), and National Experimental Cell Resource Platform (NECRP) (1 of 131). Additionally, cells were received as gifts from scientists from other universities (4 of 131). However, the authors did not report the original source of their gifted RBL cell line. Finally, the cell origin was not specified in 33 out of 131 publications. Figure 4 summarizes the cell line sources of RBL-2H3.
ATCC and BCRC recommends RBL-2H3 to be cultured in MEM that contains Earle’s Balanced Salt Solution, non-essential amino acids, 2 mM L-glutamine, 1 mM sodium pyruvate, made complete with the supplement of 15% FBS. CCTCC and JCRB recommend using the same MEM base media with slight reduction of FBS to 10%. KCLB, however recommends DMEM with 10% FBS. For institutions other than the aforementioned above, there is no information regarding the recommended media used for the culture of RBL-2H3.
Review of published methods revealed significant variation in the culture media and supplements used to cultivate RBL-2H3 cells (Table 5). However, 11 out of 131 studies did not disclose the culture conditions used. DMEM was the most used (69 of 131), followed by MEM (44 of 131), MEM Eagle-alpha modification (2 of 131), RPMI-1640 (3 of 131), and EMEM (1 of 131). For serum, media were mostly supplemented with 10% FBS (91 of 131), followed by 15% FBS (26 of 131), 20% FBS (1 of 131) 17% FBS (1 of 131) and 5% FBS (1 of 131). Other supplements reported include penicillin/streptomycin (102 of 131), L-glutamine/GlutaMAX (16 of 131), Sodium pyruvate (14 of 131) and non-essential amino acid (NEAA) (7 of 131). Two study utilized distinctive medium formulation for the culture of RBL-2H3 cells: a concoction of 70% α-MEM, 20% RPMI 1640 and 10% FBS with 2mM L-glutamine, as well as a mix of 41.5% MEM, 41.5% IMDM and 17% FBS with 25 mM HEPES and 120 μg/mL gentamicin sulphate. There is yet to be any report on the influence of various media and supplements on RBL-2H3 cells physiology and function.