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iPS cell culture supernatant as well as preparation method and application thereof

A technology of cell culture and supernatant, applied in the field of cells, can solve the problems of high cost, complicated extraction process of iPS cell extract, long cell culture cycle, etc., and achieve the effect of easy acquisition

Pending Publication Date: 2020-01-24
XIAN INT UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Studies have shown that iPS cells have the function of tissue repair. At present, the application of iPS cells basically uses iPS cell extracts, but the extraction process of iPS cell extracts is complicated, the cell culture cycle is long, the cell proliferation is slow, and the amount of cells lost by cell extraction is too large , high cost, not conducive to large-scale use

Method used

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  • iPS cell culture supernatant as well as preparation method and application thereof
  • iPS cell culture supernatant as well as preparation method and application thereof
  • iPS cell culture supernatant as well as preparation method and application thereof

Examples

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preparation example Construction

[0025] The preparation method of the iPS cell culture supernatant (iPS culture medium, iPS-CM) of the present invention comprises:

[0026] Step 1, culture of induced pluripotent stem cells (iPS): Matrigel matrigel was allowed to stand at 37°C for 1 hour, cultured with mTeSR+5×Supplement medium, inoculated with induced pluripotent stem cells, and incubated at 37°C in CO 2 Cultivate in an incubator, expand and culture for 3-7 days, so that the iPS cell concentration is 1-7×10 5 cells / mL to obtain iPS cell culture fluid;

[0027] Step 2, preparation of iPS cell culture supernatant: collect the supernatant of iPS cell culture, store it in cold storage at -20°C, concentrate by centrifugation (2-6 times of low-speed centrifugation or high-speed centrifugation), and concentrate the volume to 1% of the original volume. / 8, iPS cell culture supernatant was obtained.

[0028] It is also possible to add other effect-producing preparations to the iPS cell culture supernatant and make a...

preparation Embodiment 1

[0035] Preparation Example 1 iPS Cell Expansion and Preparation of Culture Supernatant

[0036] Culture iPS cells separately for 3 days at a cell concentration of 1×10 5cells / mL, iPS cell culture fluid was obtained, collected and concentrated three times by 2000r / min low-speed centrifugation for 10min to concentrate the volume to 1 / 8 of the original volume. 1 / 8 of the original volume, respectively, to obtain iPS-CM.

preparation Embodiment 2

[0037] Preparation Example 2 iPS Cell Expansion and Preparation of Culture Supernatant

[0038] Culture iPS cells separately for 4 days, the cell concentration is 3×10 5 cells / mL, the iPS cell culture fluid was obtained, collected and centrifuged at a low speed of 2000r / min for 10min to concentrate twice to obtain iPS-CM.

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Abstract

The invention provides an iPS cell culture supernatant as well as a preparation method and application thereof. A supernatant, which is collected through concentration at a certain rate, for iPS cellculture has remarkable growth promoting effect to human skin fibroblasts and is applied to damage repair with combination of other drugs so as to be capable of providing a long-term effective cell source by using stem cells and also specifically repairing human tissues as well as aims at maximizing the effects of the repair processes of the two of the supernatant and other drugs.

Description

technical field [0001] The invention belongs to the field of cell technology, and relates to an iPS cell culture supernatant and a preparation method and application thereof. Background technique [0002] The emergence of induced pluripotent stem cells (induced pluripotent stem cells, iPS cells) not only solves the problem of limited proliferation and differentiation ability of adult stem cells, but also avoids the ethical issues of embryonic stem cells, providing a reliable source of seed cells for regenerative medicine; Stem cells are immature cells with multilineage differentiation potential. Theoretically, stem cells have the ability to induce differentiation in the internal environment of tissue damage, that is, to realize the possibility of on-demand repair and regeneration of damaged tissues. At present, stem cell-based cell therapy is becoming a hot spot and frontier in the field of regenerative medicine research (Shi Guidong. Experimental research on iPS cell-deriv...

Claims

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Application Information

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IPC IPC(8): C12N5/074A61K35/545A61K45/06A61P17/00
CPCA61K35/545A61K45/06A61P17/00C12N5/0696A61K2300/00
Inventor 王琳刘扬王金佩杨力侠邢书娟董明清
Owner XIAN INT UNIV
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