A simple and improved method to generate human hybridomas.

T Kudo; H Saeki; T Tachibana

doi:10.1016/0022-1759(91)90317-9

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A simple and improved method to generate human hybridomas.

T Kudo·H Saeki·T Tachibana

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Abstract

By improving our previous method for production of human hybridomas, we developed a simple and remarkably efficient method for production of human hybridomas. We reformed our previous method in the following three points: (1) we added irradiated (30 Gy) myeloma cells as feeder cells to culture of fusion cells; (2) ouabain concentration in the selective medium was reduced from 2 x 10(-6) M to 1 x 10(-6) M; (3) selective medium (GIT-HAT-OL) was added to the culture after overnight cultivation of fusion cells. Consequently, we obtained higher fusion frequency (1/700 vs. 1/5500) compared with our previous method. By our present method, only so small number of human B cells (EBV-LCL) is required, so that the time necessary to establish human hybridomas is reduced.

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Cell Fusion Cells, Cultured Clone Cells Culture Media Humans Hybrid Cells Hybridomas Immunologic Techniques In Vitro Techniques Rheumatoid Factor Journal Article

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T Kudo, H Saeki, T Tachibana. 1991-12-15. A simple and improved method to generate human hybridomas.. https://doi.org/10.1016/0022-1759(91)90317-9

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