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A target regulation system, kit and application for somatic cell dedifferentiation

A technology for nerve cells and fibroblasts, which is applied in the field of target regulation systems for somatic cell dedifferentiation, can solve the problems of unclear components of the transdifferentiation system, difficult to repeat experiments, and unstable effects, and achieve easy quality control and scale. The effect of production, clear target and efficiency improvement

Active Publication Date: 2021-07-09
YUNNAN JICI INSITUTE FOR REGENERATIVE MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, Schwann cells (THOMA EC, et al, 2014), nerve cells (HU W, et al, 2015), pancreatic islet cells (Sheng The results of Ding, et al, 2015) have been reported successively, but there are still some problems in the above-mentioned studies, such as difficulty in repeating the experiment, low efficiency, unclear composition of the transdifferentiation system, and unstable effect.

Method used

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  • A target regulation system, kit and application for somatic cell dedifferentiation
  • A target regulation system, kit and application for somatic cell dedifferentiation
  • A target regulation system, kit and application for somatic cell dedifferentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Target Regulatory System Used to Prepare Osteoblasts

[0072] 1. Isolation of skin fibroblasts

[0073] 1.1 Obtain a skin tissue block with a diameter of about 1 cm from the donor, separate the skin fibroblasts by the adherent method, and culture the separated cells in the basal culture medium: 10% fetal bovine serum (Hyclone) + 100U / ml penicillin (Sigma) + 100 μg / ml streptomycin (Sigma) + high glucose DMDM.

[0074] 1.2 Subsequent passage of cells, a large number of expansion, the number of cell generation is between the 6th passage and the 12th passage, which is used to induce the transdifferentiation into osteoblasts. The day before initiation of differentiation (Day-1), inoculate cell density at 1-2.5×10 4 / cm 2 Cultured at 37°C, 5% CO 2 in the incubator.

[0075] 2. Reprogramming of skin fibroblasts: the first stage

[0076] After the treatment in the first step above, completely replace it with the induction medium of the first stage for cell cult...

Embodiment 2

[0081] Example 2 Target Regulatory System Used to Prepare Osteoblasts

[0082] 1. Isolation of skin fibroblasts

[0083] 1.1 Obtain a skin tissue block with a diameter of about 1 cm from the donor, separate the skin fibroblasts by the adherent method, and culture the separated cells in the basal culture medium: 10% fetal bovine serum (Hyclone) + 100U / ml penicillin (Sigma) + 100 μg / ml streptomycin (Sigma) + high glucose DMDM.

[0084]1.2 Subsequent passage of cells, a large number of expansion, the number of cell generation is between the 6th passage and the 12th passage, which is used to induce the transdifferentiation into osteoblasts. The day before initiation of differentiation (Day-1), inoculate cell density at 1-2.5×10 4 / cm 2 Cultured at 37°C, 5% CO 2 in the incubator.

[0085] 2. Activation of skin fibroblasts

[0086] 2.1 When starting transdifferentiation (Day0), completely replace the basal culture medium with the first-stage culture medium, and culture the ce...

Embodiment 3

[0092] Example 3 Target Regulatory System Used to Prepare Chondrocytes

[0093] 1. Isolation of skin fibroblasts

[0094] 1.1 Obtain a skin tissue block with a diameter of about 1 cm from the donor, separate the skin fibroblasts by the adherent method, and culture the separated cells in the basal culture medium: 10% fetal bovine serum (Hyclone) + 100U / ml penicillin (Sigma) + 100 μg / ml streptomycin (Sigma) + high glucose DMDM.

[0095] 1.2 Subsequent passage of cells to a large number of expansion, the number of cell generations between the 6th and 12th passages is used for the induction of transdifferentiation into chondrocytes. The day before initiation of differentiation (Day-1), inoculate cell density at 1-2.5×10 4 / cm 2 Cultured at 37°C, 5% CO 2 in the incubator.

[0096] 2. Reprogramming of skin fibroblasts

[0097] After the treatment in the second step above, completely replace it with the second-stage culture medium for cell culture. The culture time is 6-12 day...

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Abstract

The invention discloses a target regulation system, kit and application for somatic cell dedifferentiation. The target regulation system regulates somatic cells as follows: inhibiting PKC activity, and inhibiting DNMT activity and / or inhibiting HMT activity and / or inhibiting histone demethylase activity; the key targets for corresponding regulation include PKC, and DNMT and / or HMT and / or histone demethylase; the somatic cells are differentiated cells. The target regulation system provided by the present invention can promote the efficiency of small molecules inducing the transdifferentiation of somatic cells into different types of functional target cells, and has clear targets, clear components, mature and stable, easy quality control and large-scale production.

Description

technical field [0001] The invention relates to the fields of cell biology, tissue engineering and regenerative medicine, in particular to a target regulation system, kit and application for somatic cell dedifferentiation. Background technique [0002] Stem cells are the source of the human body and its various tissue cells. They are of great significance in the research of cell therapy of human diseases, regenerative medicine, tissue engineering, drug development, anti-aging, beauty and many other fields. However, due to the ethical issues and potential safety hazards of embryonic stem cells; the scarcity of various types of adult stem cells, limited sources, difficulty in enrichment, complicated acquisition process, restrictions on the health status of donors, easy aging of cells, and the inability to expand a large number of generations, etc., Therefore, they have not been widely used. A variety of differentiated cells such as skin fibroblasts have the advantages of abun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077
CPCC12N2501/01C12N2501/415C12N2501/15C12N2501/72C12N5/0656C12N2501/727
Inventor 胡敏李燕皎
Owner YUNNAN JICI INSITUTE FOR REGENERATIVE MEDICINE CO LTD
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