血清飢餓培養の実現を目指した, ハイブリドーマ細胞への細胞死耐性能の付与 [in Japanese] Establishing an Apoptosis-Resistant Hybridoma Cell Line for Usage in Serum Starvation Culture [in Japanese]
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ハイブリドーマ細胞の血清飢餓培養には抗体生産能の上昇などの利点があるが, 急速な細胞死 (アポトーシス) を生じ, 実用的でない.そこで細胞にアポトーシス抑制遺伝子<I>bcl-2</I>を導入し, 更に血清飢餓でスクリーニングして血清飢餓耐性株を樹立した.この株はスクリーニング前の1.4倍のBcl-2蛋白を発現し, 野性株が2日で死滅する0.2%血清培地中で4日の培養が可能で, 通常の培養の3倍の比抗体生産速度を示した.
A serum starvation culture of hybridoma cells is desirable because serum starvation stops cell cycling at the G<SUB>1</SUB> phase, where hybridoma cells produce more antibody in the other phases, However, serum starvation induces apoptosis, a type of cell death, forcing the culture to die out. In this research, the authors aimed at inhibition of cell death during serum starvation culture. A hybridoma cell line <I>bcl-2</I> was transfected with <I>bcl-2</I>, an apoptosis suppressing gene, and named 2E3-<I>bcl 2</I>. Then the cells were screened in serum starvation culture to obtain 2E3-<I>bcl 2</I>-LS. The expression level of Bcl-2 protein in 2E3-<I>bcl 2</I>-LS was higher by 40% than that in 2E3-<I>bcl-2</I>. In serum starvation culture, 2E3-<I>bcl 2</I>-LS survived longer by two days and produced twice as much antibody as the wild type.
- Chemical engineering
Chemical engineering 24(1), 161-164, 1998-01-10
The Society of Chemical Engineers, Japan