Method for culturing animal cells in collagen drops on a support

Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Method of culturing encapsulated cells

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435 29, 435177, 435182, 435395, 435404, C12N 500, C12N 102, C12N 1104, C12Q 102

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057121613

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BRIEF SUMMARY
TECHNICAL FIELD

The present invention relates to a method for culturing animal cells under embedded conditions, and in particular, it relates to a method for culturing animal cells under embedded conditions, which method is applicable to a method for testing the sensitivity to anticancer agents or a method for testing co-culture.


BACKGROUND ART

As to conventional tests for the sensitivity to anticancer or carcinostatic agents, they may be carried out using subcultured cancer cells, but methods of evaluating effects of anticancer agents upon respective individuals by utilizing so-called primary culture in which samples taken from a living body are directly cultured are widely employed.
However, when using a sample taken from a living body, only a small amount of cancer cells can be obtained. In addition, the sample contains normal cells other than cancer cells of interest and further contains many other components.
Tests for the sensitivity to anticancer agents (anticancer agent sensitivity tests) are conventionally carried out, using different anticancer agents alone or in various combinations, or changing the dose of anticancer agent to various ones, thus discovering optimum conditions. For this reason, a sample taken from a living body is divided under respective test conditions, and only an aliquot of the sample is used for each test. However, as mentioned above, if the total amount of cancer cells in the sample taken from the living body is small, the amount of cancer cells as separated for use in each test become very slight. Consequently, the accuracy of the test is lowered or a sufficient amount of samples to provide to necessary test items cannot be obtained. In addition, the proliferation of cancer cells may be inhibited even when cultured under conditions where the cancer cells are not in contact with anticancer agents, because of mass proliferation of cells other than the cancer cells, such as fibroblasts. Furthermore, the cancer cells may be difficult to distinguish from other cells when evaluating culture effect. This renders it impossible to accurately compare proliferation states of the cancer cells when in contact with the anticancer agent and when not in contact therewith.
Thus, various methods for increasing the accuracy of anticancer agent sensitivity tests have been proposed. An example of the methods includes embedding a sample containing cancer cells in a collagen gel for culturing. Because the proliferation of cancer cells is quite favorable in collagen gels, the comparison between the results of adding and not adding an anticancer agent in question is facilitated. There is another method in which the number of colonies formed upon proliferation of cancer cells is counted utilizing image analysis technique. This method is very favorable for using a sample which is taken from a living body and contains a mixture of cancer cells and fibroblasts, because this method allows a morphological distinction in a collagen gel between colonies of cancer cells formed into in-vivo-like morphology and colonies formed by the proliferation of fibroblasts contained together with the cancer cells in the sample taken from a living body. This technique, in which the collagen gel-embedded culture and the image analysis are combined, was earlier invented by the inventors of the present application and is disclosed in the specification of Japanese Patent Application No. Heisei 2-267343(Japanese Official Patent Provisional Publication (Kokai) No. Heisei 3-285696).
However, even the above-mentioned conventional technique is unsatisfactory in respect to culture technique in order to increase the accuracy of anticancer agent sensitivity tests. Increase in accuracy of anticancer agent sensitivity tests requires seeding cancer cells at a appropriate cell density even in a small amount of cancer cells and requires culturing under an environment as favorable as possible to the proliferation of cancer cells.
In a culture method in which a collagen solution containing dispersed cancer cells is poured into a cultu

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Chem. Abstracts, vol. 115, No. 15, Abstract No. 149651p Aizawa et al. "Usefulness of Collagen-Gel Matrix Culture . . . ".
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