Method for sorting cardiac fibroblast CD90<+> subsets of mice and application of cardiac fibroblast CD90<+> subsets

A technique for cardiac fibroblasts and fibroblasts, applied in the field of sorting rat cardiac fibroblasts CD90+ subsets, which can solve the problems of cell proliferation and collagen synthesis differences

Inactive Publication Date: 2018-04-06
XINXIANG MEDICAL UNIV
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Problems solved by technology

[0006] It has been found that CD90 (Thy1) is also expressed in other tissue fibroblasts, and the fibroblasts of rat lung, heart, human orbit and reproductive tract can be divided into Thy1 - Group and Thy1 + groups, there were significant differences in cell proliferation and collagen synthesis, but CD90 + The role of CFs in the differentiation of cardiomyocytes and the repair of myocardial damage after cell transplantation have not been reported yet.

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  • Method for sorting cardiac fibroblast CD90&lt;+&gt; subsets of mice and application of cardiac fibroblast CD90&lt;+&gt; subsets
  • Method for sorting cardiac fibroblast CD90&lt;+&gt; subsets of mice and application of cardiac fibroblast CD90&lt;+&gt; subsets
  • Method for sorting cardiac fibroblast CD90&lt;+&gt; subsets of mice and application of cardiac fibroblast CD90&lt;+&gt; subsets

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[0029] Materials and Methods

[0030] 1.1 Experimental animals 80 adult SD rats, male or female, weighing 205.6 ± 0.56 g, were provided by the Experimental Animal Center of Xinxiang Medical College (animal qualification certificate number: SCXK (Yu) 2010-0002).

[0031] Cell line Rat cardiac fibroblasts (R6300, purchased from ScienCell, USA)

[0032] 1.3 Recovery and expansion of cell lines

[0033] Thaw the cell line cryopreservation tube quickly in a 37°C water bath, centrifuge at 1000 rpm for 3 min, discard the supernatant, and inoculate to a temperature of 75 m 2 In culture flasks, the medium was changed after 24 h. Afterwards, the medium was changed every 48 h, and when the cells grew to a confluent state, the cells were digested with trypsin-EDTA digestion solution and counted at 1×10 5 cells / mL in a culture flask at 37°C, 5% CO 2 Expand the culture in the incubator.

[0034] Cell slides were fixed with 4% paraformaldehyde, rinsed with PBS, and washed with 3% H 2 o...

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Abstract

The invention belongs to the animal cell culture techniques and relates to a method for sorting cardiac fibroblast CD90<+> subsets of mice and application of the cardiac fibroblast CD90<+> subsets. The method comprises the following steps: carrying out recovery and enlarged culture on mice cardiac fibroblast cell strains; sorting cardiac fibroblast CD90<+> subsets by virtue of a PE-marked CD90 antibody; collecting the sorted cardiac fibroblast CD90<+> subsets, carrying out centrifugation to remove supernatant so as to obtain precipitates, inoculating the precipitates into a culture bottle, carrying out enlarged culture, so as to finish the sorting of the cardiac fibroblast CD90<+> subsets of the mice. The sorted cardiac fibroblast CD90<+> subsets are taken as preponderant subsets for sorting cardiac fibroblasts and are used for triggering the myocardium repair potential of the cardiac fibroblasts, so that the experimental basis and theoretical support are provided for the efficient utilization of the cardiac fibroblasts.

Description

technical field [0001] The invention belongs to animal cell culture technology, and relates to a method for sorting rat heart fibroblast CD90 + Subgroup methods and their applications. Background technique [0002] Ischemic heart disease has become the number one killer threatening human health and life, how to prevent and treat ischemic heart disease has attracted great attention. At present, repairing damaged myocardium is mainly to replenish or replace necrotic cardiomyocytes while preventing myocardial fibrosis. In recent years, with the gradual deepening of stem cell research, cell transplantation is considered to be a regenerative medicine therapy with great clinical application prospects for the treatment of ischemic heart disease. Then, in addition to the directional repair of stem cells, can other cells with strong proliferation ability in myocardial tissue be mobilized to induce differentiation into new cardiomyocytes, so that heart function can be effectively im...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077A61K35/34A61K35/33A61P9/10
CPCA61K35/33A61K35/34C12N5/0656C12N5/0657C12N2501/06
Inventor 常玉巧李辞霞郭志坤王现伟贾阳阳郭永龙张俊悦
Owner XINXIANG MEDICAL UNIV
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