Cell-Based Screening of Traditional Chinese Medicines for Proliferation Enhancers of Mouse Embryonic Stem Cells

摘要

A high-throughput cell-based method was developed for screening traditional Chinese herbal medicines (TCHMs) for potential stem cell growth promoters. Mouse embryonic stem (mES) cells expressing enhanced green fluorescent protein (EGFP) were cultured in growth media supplemented with various TCHM extracts. The dosage-dependent effects of TCHM extracts on cell growth, including proliferation and cytotoxicity, were assessed via EGFP fluorescence measurement. Seven TCHMs were investigated, and among them Panax noto-ginseng (PN), Rhizoma Atractylodis macrocephalae, Rhizoma chuanxiong, and Ganoderma lucidum spores (GLS) showed potential to improve mES cell proliferation. Eleven mixtures of these four TCHMs were then studied, and the results showed that the mixture of PN and GLS had the strongest growth promoting effect, increasing the specific growth rate of mES cells by 29.5% at a low dosage of 0.01% (wt/vol) PN/GLS (P<0.01) and 34.2% at 0.1% (wt/vol) PN/GLS (P < 0.05) compared to the control. The growth promoting effect of PNI GLS was further confirmed with ES cells cultured in spinner flasks. A 29.3-fold increase in the total cell number was achieved in the medium supplemented with 0.01 % PN/GLS after 5 days, while the control culture only gave a 16.8-fold increase. This cell-based screening method thus can provide an efficient and high-throughput way to explore potential stem cell growth promoters from TCHMs.