Shanghai Innovative Chang'an Biological Technology Co., Ltd.

Lingyan Zhu

Jing Wang

Xiaoyan Zhang

Jianqing Xu

The present invention relates to a method for inducing amplification of human type I NKT cells in vitro using a 'specific stimulant+staged cytokine' mode, which consists of two culture stages, wherein the first culture stage focuses on specific amplification of the number of the type I NKT cells, in which a specific stimulant &#x3b1;-GalCer is used to advantageously amplify the type I NKT cells and &#x3b1;-GalCer-loaded CD1d-expressing cells are used to stimulate continuous proliferation of the type I NKT cells while adding cytokines IL-2 and IL-7 to assist growth of the type I NKT cells; and the second culture stage is to synchronously perform amplification of the number of the type I NKT cells and guide directed function differentiation, in which CD1d-expressing cells incubated with &#x3b1;-GalCer continue to stimulate proliferation of the type I NKT cells while adding IL-2, IL-7 and IL-15 to assist amplification of the type I NKT cells and guide differentiation, and IL-12 is added to the culture system 1-2 days before the end of culture to guide further directed differentiation of the type I NKT cells and enhance their killing activity. The method of the present invention is simple to operate, can greatly amplify the type I NKT cells in vitro while improving the killing activity of the amplification products, and is suitable for large-scale production.

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Shanghai, China

Lingyan Zhu