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Cell culture composition used for primary culture of tumor cells and application thereof

A technology for cell culture and tumor cells, applied in the biological field, can solve problems such as low success rate, achieve stable passage, improve primary culture success rate, and high purity

Inactive Publication Date: 2014-08-06
高全立
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the success rate of culturing primary cancer cells in serum-free or low-serum medium is still very low, most of which do not exceed 20%, and the culture time usually needs to be more than 1 month

Method used

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  • Cell culture composition used for primary culture of tumor cells and application thereof
  • Cell culture composition used for primary culture of tumor cells and application thereof
  • Cell culture composition used for primary culture of tumor cells and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Embodiment 1: cell culture composition 1 and preparation method thereof

[0093] Cell Culture Composition 1:

[0094] Invitrogen's Defined Keratinocyte-SFM: 500ml;

[0095] Invitrogen's B27supplement50×: 10ml;

[0096] Total: 510ml.

[0097] Preparation method: mix the above keratinocyte serum-free medium with B-27 to obtain cell culture composition 1.

Embodiment 2

[0098] Embodiment 2: cell culture composition 2 and preparation method thereof

[0099] Cell Culture Composition 2:

[0100] Sigma's Keratinocyte Medium II: 500ml;

[0101] Invitrogen's B27supplement50×: 2ml;

[0102] Total: 502ml.

[0103] Preparation method: mix the above keratinocyte serum-free medium with B-27 to obtain cell culture composition 2.

Embodiment 3

[0104] Embodiment 3: cell culture composition 3 and preparation method thereof

[0105] Cell Culture Composition 3:

[0106] Invitrogen's keratinocyte-SFM: 500ml;

[0107] Invitrogen's B27 supplement without vitamin A 50×: 10ml;

[0108] Epidermal growth factor: 2550ng;

[0109] Total: 510ml.

[0110] Preparation method: mix the above keratinocyte serum-free medium with B-27 additive and epidermal growth factor to obtain cell culture composition 3.

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Abstract

The invention provides a cell culture composition used for primary culture of tumor cells and application thereof. According to application of the cell culture composition in primary culture of tumor cells from epitheliums, the cell culture composition comprises a serum-free horn cell medium and a serum-free additive for culture of nerve cells, and does not contain serum. The cell culture composition provided by the invention can realize successful in vitro primary culture of the tumor cells from epitheliums, substantially increases the success rate of primary culture of the tumor cells from epitheliums and can inhibit growth of stromal cells like fibroblasts and endothelial cells in cancer tissue; for example, primary culture of the tumor cells from epitheliums succeeds within 10 d by using the cell culture composition provided by the invention, and the success rate of culture is mostly more than 80%; the cultured tumor cells can be stably passed to more than 8 generations, and a cell line can be successfully established.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cell culture composition for primary culture of tumor cells, its application, and a primary culture method. Background technique [0002] In the current tumor cell culture technology, the most commonly used technology for primary culture and line establishment of tumor cells is to use a basal medium supplemented with 10% fetal calf serum (FCS for short) to cultivate tumor cells. However, the current application of this tumor cell culture technology to cultivate primary tumor cells has a very low success rate. For example, the success rate of primary lung cancer and prostate cancer cells cultured with 10% FCS+RPMI1640 is lower than 10%, and the success rate of subculture and line establishment is even lower. Therefore, currently established cell lines are mostly used in tumor research. Since most of the established cell lines have been cultured and passaged in vitro fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 高全立
Owner 高全立
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