Large-scale, high-density freezing of hybridomas and its application to high-density culture |
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Authors: | Ninomiya N Shirahata S Murakami H Sugahara T |
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Institution: | Graduate School of Genetic Resources Technology, and Department of Food Science and Technology, Faculty of Agriculture, Kyushu University 46-09, 6-10-1 Hakozaki, Higashi-Ku, Fukuoka 812 Japan. |
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Abstract: | Large-scale, high-density freezing of hybridomas was studied to apply frozen cells to start high-density culture. We showed here that hybridomas can be frozen at 1.5 x 10(8) cells/mL, without decrement in viability and proliferating activity. Blood transporting bags were used for large-scale freezing to store 25 mL of cell suspension with a cell density, 1.5 x 10(8)/mL. The number of cells stored in a bag (3.0 x 10(9) cells) was enough to start a high-density culture at a 10 times higher cell density (6.0 x 10(6) cells/mL) than normal inoculation, and the cells proliferated to 10(7) cells/mL within 2 days. These results indicate that the large-scale freezing method is useful for large-scale culture of mammalian cells. |
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Keywords: | high-density freezing high-density culture hybridoma |
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