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Technology for regulating and controlling Jak-Stat pathway to differentiate, dedifferentiate and rejuvenate cells, and application of technology

A cell differentiation and de-differentiation technology, applied in animal cells, cell culture active agents, genetically modified cells, etc., can solve the problems of tumorigenesis, genetic variation, unfavorable clinical application, etc.

Pending Publication Date: 2019-11-08
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

However, the induction efficiency of iPS is low, and involves the introduction of exogenous genes, and genetic variation often occurs, which is not conducive to clinical application
Genetic modification has recently been proposed as a rejuvenation method, but this method still has the risk of off-target and tumorigenesis

Method used

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  • Technology for regulating and controlling Jak-Stat pathway to differentiate, dedifferentiate and rejuvenate cells, and application of technology
  • Technology for regulating and controlling Jak-Stat pathway to differentiate, dedifferentiate and rejuvenate cells, and application of technology
  • Technology for regulating and controlling Jak-Stat pathway to differentiate, dedifferentiate and rejuvenate cells, and application of technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1, the acquisition and identification of repairing skin fibroblasts

[0102] 1. Human skin fibroblasts were inoculated into 6-well plates and cultured in skin fibroblast culture medium for 24 hours;

[0103] 2. Replace the cell culture medium with the rFib induction medium containing the small molecule cocktail combination Mix V, and change the medium every 2 days;

[0104] 3. After culturing for 5 days with the rFib induction medium containing the small molecule cocktail combination Mix V, replace the culture medium with the rFib induction medium containing the small molecule cocktail combination Mix P, and change the medium once every 2 days;

[0105] 4. After culturing for 7 days with the rFib induction medium containing the small molecule cocktail combination Mix P, the medium was replaced with HG-DMEM supplemented with 10% FBS, 10ng / mL bFGF, 100ng / mL PDGF-AB and 10ng / mL BMP4 ; Or add 10% FBS to HG-DMEM; Or use rFib medium for culture; Cells are treated for...

Embodiment 2

[0152] Example 2 rFib cells have the ability to repair bone defects

[0153] Under the approval of the ethics committee, NOD / SCID mice aged 8 to 10 weeks and weighing 20 to 24 g were used to create a femoral defect model, and 5 animals were used in each group. The modeling method was as follows: under pentobarbital sodium anesthesia, cut The skin and subcutaneous tissue were opened, and a sufficient mid-femur was exposed by blunt dissection between the rectus femoris and semitendinosus. The operation was performed at the proximal junction of the center of the right femur. A continuous bone defect of 4mm×1mm was constructed surgically. Skin fibroblasts (Fib), bone marrow mesenchymal stem cells (bMSC) and rFib were stained with Hoechst 33342 (Thermo, NucBlue live cell), then mixed with Matrigel and transplanted into the defect site with 5×105 cells / mouse.

[0154] Twenty-eight days after transplantation, the mice were sacrificed by injecting a lethal dose of sodium pentobarbit...

Embodiment 3

[0161] Example 3, rFib cells have the ability to repair cartilage defects

[0162] Articular cartilage defect model and cell transplantation

[0163] NOD / SCID mice weighing 20-24 g at the age of 8 to 10 weeks were used. A modified articular cartilage model was used to evaluate the efficacy of rFib (Cheng et al., 2014). Articular cartilage defects (1.5 mm x 1 mm) were constructed in the trochlear groove of the distal femur with a biopsy punch. cells ((2.5×10 5 Each in 35 μl Matrigel) was labeled with Hoechst 33342 and implanted into the defect site. Matrigel implants without cells were used as controls.

[0164] Figure 7 .In vivo cartilage repair experiment

[0165] A-B. Safranin Fast Green staining of cartilage tissue samples and 10 μm sections. In safranin fast green staining, red is cartilage and green is bone tissue. Both young rFib (39 years old) and bMSCs (31 years old) can repair cartilage defects. Although old bMSCs (62 years old) can generate new tissue, they ...

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Abstract

The present invention belongs to the technical field of cell biology and particularly relates to a technology for regulating and controlling a Jak-Stat pathway to differentiate, dedifferentiate and rejuvenate cells, and an application of the technology. Rejuvenated cell products and / or cell products of different lineage types are obtained by small molecule compound combination, cytokine combination or recombinant protein combination, a gene editing technology and a transgenic technology to quantitatively and / or regularly regulate and control gene or protein targets of the Jak-Stat signaling pathways in cells. The provided technology for regulating and controlling the Jak-Stat signal pathway, and the product / derivatives of the cell product can be used for re-programming cells, tissues, organs and organisms, construct tissue engineering materials, repair damages of tissues and organs of mammals and the aged and degenerated tissues and organs, delay or reverse aging processes of the cells, tissues, organs and organisms, and can be used for immunoregulation of the cells, tissues, organs and organisms.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and relates to a technique for regulating the Jak-Stat pathway to make cells differentiate, dedifferentiate and rejuvenate and its application, especially to regulate the JAK-STAT signal pathway to regulate cell differentiation, dedifferentiation and transdifferentiation , rejuvenation, aging, apoptosis and methods for prolonging the life of the body and the preparation and application of cell products. Background technique [0002] Stem cells are considered the "holy grail" of regenerative medicine and anti-aging. When an individual begins to age, the stem cells in the body will also age, leading to malignant transformation or degeneration of multiple organs including bone, cartilage, heart, muscle, brain, skin, pancreas, liver, kidney, and gastrointestinal tract. Improper immune system function is also associated with aging. In fact, chronic inflammation of tissues and organs is the caus...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0775C12N5/10C12N15/867C12N15/90A61K35/33A61K35/28A61P39/06A61P37/02A61K8/99A61Q19/08A61L27/38A23L33/00
CPCC12N5/0656C12N5/0663C12N15/86C12N15/907C12N9/22A61K35/33A61K35/28A61P39/06A61P37/02A61K8/99A61Q19/08A61L27/3804A61L27/3834A23L33/00C12N2501/40C12N2501/415C12N2501/42C12N2501/065C12N2501/15C12N2501/01C12N2510/00C12N2800/107C12N2810/10C12N2740/15043A61L2300/412A23V2002/00A23V2200/30A61P19/10A61P17/02A61P9/10A61K8/64C12N5/0619C12N5/0662C12N2506/1307C12N2501/115C12N2501/135C12N2501/155C12N2501/727C12N2501/385A23L33/10C12N5/0668C12N2501/999
Inventor 胡敏李燕皎
Owner YUNNAN JICI INSITUTE FOR REGENERATIVE MEDICINE CO LTD
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