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Rapid extraction method of fibroblast and application

A technology of fibroblasts and extraction methods, which is applied in the field of rapid extraction of fibroblasts, and can solve the problem of not being able to obtain tumor-related fibroblasts

Inactive Publication Date: 2016-10-26
GUANGXI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the existing method of extracting tumor-associated fibroblasts, a large number of tumor-associated fibroblasts cannot be obtained, which has a great impact on the study of its mechanism and function

Method used

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  • Rapid extraction method of fibroblast and application
  • Rapid extraction method of fibroblast and application
  • Rapid extraction method of fibroblast and application

Examples

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Embodiment Construction

[0028] All features disclosed in this specification, or steps in all methods or processes disclosed, may be combined in any manner, except for mutually exclusive features and / or steps.

[0029] Any feature disclosed in this specification (including any appended claims, abstract), unless otherwise stated, may be replaced by alternative features that are equivalent or serve a similar purpose. That is, unless expressly stated otherwise, each feature is one example only of a series of equivalent or similar features. The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0030] 1. Materials and reagents used in the examples

[0031] The ophthalmic scissors are products of Dadu Instrument Medical Co., Ltd., and the model is YYJ-PT>10cm curved scissors. The end of the ophthalmic scissors is 0.4 mm wide, pointed but not sharp.

[0032] Human liver cancer HepG2 cells (ATCC, product number HB-8065), the public c...

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Abstract

The invention discloses a rapid extraction method of fibroblast. The method comprises the following steps that 1, isolated tumor tissue is taken and cut into pieces, and a collagenase solution is added for digestion; 2, the mixture is transferred into a centrifugal tube for oscillation, incubation and centrifuging, and sediments are taken to be placed into a cell incubator for culture; 3, the collagenase solution is added for digestion, and the step 2 is executed again to obtain tumor tissue of the loose organization structure obtained after alternate digestion and culture; 4, a pancreatin solution is added, fetal cattle serum is added after normal temperature standing for terminating digestion, suspended solids are poured away, and attaching-wall tumor single cells obtained after trypsin digestion are obtained; 5, a DMEM complete medium is added for subculture, tumor-associated fibroblast is obtained. The invention further provides application of the extraction method. The fibroblast obtained through the extraction method is high in purity, the fibroblast highly expresses FAP, the expression rate is 99.2%, and the method is suitable for medicine popularization.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a rapid extraction method and application of fibroblasts. Background technique [0002] Cancer-associated fibroblasts (CAFs) are the most important cellular components in the tumor stroma and play a crucial role in the occurrence and development of tumors. CAF can recruit endothelial progenitor cells to local tumors to promote tumor angiogenesis and tumor growth. In addition to expressing vimentin, a marker of fibroblasts, these tumor-associated fibroblasts also specifically express FAP and α-SMA, but not CK and desmin. At the genetic level, it is significantly different from normal fibroblasts (NF). [0003] Tumor-associated fibroblasts are currently one of the research hotspots, and many mechanisms are still controversial, but it is generally believed that the cells are inseparable from the occurrence, development, metastasis and invasion of tumors. Tumor-associated fi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 赵永祥卢小玲周素芳黄盈盈
Owner GUANGXI MEDICAL UNIVERSITY
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