Chemical induction method for reprogramming dermal fibroblasts into endothelial progenitor cells

A technology of fibroblasts and endothelial progenitor cells, applied in the field of cell engineering, can solve the problems of easy side effects, complex technical methods, high cost and so on

Pending Publication Date: 2022-03-25
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Usually, multiple small molecule compounds are combined for induction in order to obtain a good induction effect. There are few research reports on the use of a single small molecule compound for induction, because the current research status shows that chemical induction with a single small molecule is prone to side effects. A good induction effect needs to pay a higher cost and adopt more complicated technical methods

Method used

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  • Chemical induction method for reprogramming dermal fibroblasts into endothelial progenitor cells
  • Chemical induction method for reprogramming dermal fibroblasts into endothelial progenitor cells
  • Chemical induction method for reprogramming dermal fibroblasts into endothelial progenitor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Isolation and culture of fibroblasts from mouse skin:

[0040] ① Newborn mice were killed one day after birth, after being disinfected with 75% alcohol, they were rinsed with 0.90% normal saline, and the skin tissue of the whole body was peeled off;

[0041] ②Remove subcutaneous fat and blood vessels, cut into pieces, and digest with 0.2% type I collagenase for 1 hour;

[0042] ③Filter with a 70um cell sieve to obtain a centrifuged suspension of the cells and inoculate them into a petri dish. CO 2 Incubate in an incubator, and subculture every 2-3 days;

[0043] (2) Compound FSK induces fibroblasts;

[0044] ① Add DMSO to FSK to dissolve, configure it as a 10 μmol / L solution, add it to the fibroblasts obtained in step (1), and continue to place it in 37°C CO 2 Incubation was carried out in an incubator for 7 days, and the cell culture medium was replaced every two days to obtain endothelial progenitor cells.

Embodiment 2

[0046] (1) Isolation and culture of fibroblasts from mouse skin:

[0047] ① Newborn mice were killed one day after birth, after being disinfected with 75% alcohol, they were rinsed with normal saline, and the skin tissue of the whole body was peeled off;

[0048] ②Remove subcutaneous fat and blood vessels, cut into pieces, and digest with 0.1% type I collagenase for 2 hours;

[0049]③Use a 70 μm cell sieve to filter, obtain cells for centrifugation suspension, and inoculate them in a culture dish. CO 2 Incubate in an incubator, and subculture every 2-3 days;

[0050] (2) Compound FSK induces fibroblasts;

[0051] ① Add DMSO to FSK to dissolve, configure it as a 5 μmol / L solution, add it to the fibroblasts obtained in step (1), and continue to place it in 37°C CO 2 Incubation was carried out in an incubator for 10 days, and the cell culture medium was changed every two days to obtain endothelial progenitor cells.

Embodiment 3

[0053] (1) Isolation and culture of fibroblasts from mouse skin:

[0054] ① Newborn mice were killed one day after birth, after being disinfected with 75% alcohol, they were rinsed with normal saline, and the skin tissue of the whole body was peeled off;

[0055] ②Remove subcutaneous fat and blood vessels, cut into pieces, and digest with 0.3% type I collagenase for 50 minutes;

[0056] ③Filter with a 70um cell sieve to obtain a centrifuged suspension of the cells and inoculate them into a petri dish. CO 2 Incubate in an incubator, and subculture every 2-3 days;

[0057] (2) Compound FSK induces fibroblasts;

[0058] ① Add DMSO to FSK to dissolve, configure it as a 15 μmol / L solution, add it to the fibroblasts obtained in step (1), and continue to place it in 37°C CO 2 Incubate in an incubator for 1 day to obtain endothelial progenitor cells.

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Abstract

The invention relates to the technical field of cell engineering, and provides a chemical induction method for reprogramming dermal fibroblasts into endothelial progenitor cells. The appropriate small molecule compound FSK is screened out to induce the fibroblasts, obvious expression of specific markers CD31 and CD34 of the endothelial progenitor cells exists in the induced cells, it is indicated that the FSK successfully achieves the technology that the dermal fibroblasts are induced into the endothelial progenitor cells through single small molecules, the technical scheme is simple, the cost is low, no side reaction occurs, and the method is suitable for large-scale popularization and application. Clinical application is facilitated, and practical significance and clinical value are achieved.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a chemical induction method for reprogramming dermal fibroblasts into endothelial progenitor cells. Background technique [0002] Angiogenesis has always been an important step in regenerative medicine, which covers the regeneration of blood vessels in a variety of diseases, including myocardial ischemia repair, liver transplantation, microcirculation reconstruction, regeneration of skin accessory vessels, etc. In order to promote the regeneration of blood vessels in various organs, people have tried various methods such as genetic modification, stem cells, and organ transplantation. Follow up question. After the skin tissue is traumatized, the outcome is mostly scar repair, accompanied by tissue ischemia and angiogenesis disorder, so the later repair and functional recovery will be seriously affected. [0003] Yamanaka, a Japanese scholar who won the Nobel Prize in Me...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0692C12N2506/1307C12N2501/999Y02A50/30
Inventor 仲苓芝李美蓉付小兵侯倩
Owner GENERAL HOSPITAL OF PLA
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