Method of inducing dedifferentiation of somatic cells with small molecules to prepare rejuvenated mesenchymal stem cells and uses thereof
AssigneeShenzhen Alpha Biopharmaceutical Co. Ltd.
InventorsMin Hu, Yanjiao Li, Junyuan Hu
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Abstract
A preparation method and applications of rejuvenated and regenerative fibroblasts, where the rejuvenated and regenerative fibroblasts are prepared from normal fibroblasts by inhibiting the JAK-STAT signaling pathway. The rejuvenated and regenerative fibroblasts are prepared by treating the target cells with a small molecular combination, a cytokine combination or a recombinant protein combination. This application further provides an application of the rejuvenated and regenerative fibroblasts in the reprogramming or rejuvenation of cells, tissues, organs and organisms.
Claims12
A method of preparing rejuvenated and regenerative fibroblasts, comprising:
- treating fibroblasts sequentially with a first composition and a second composition;
- wherein the first composition comprises high glucose DMEM (HG-DMEM) supplemented with 10% fetal bovine serum (FBS) and a first small molecule mixture consisting of WNT/β-catenin agonist CHIR99021, histone deacetylase (HDAC) inhibitor valproic acid (VPA), cyclic adenosine monophosphate (cAMP) agonist Forskolin, and transforming growth factor (TGF)-β inhibitor Repsox;
- and
- the second composition comprises HG-DMEM supplemented with 10% FBS and a second small molecule mixture consisting of HDAC inhibitor VPA, TGF-β inhibitor Repsox, WNT/β-catenin agonist CHIR99021, cAMP agonist Forskolin, retinoic acid receptor (RAR) agonists AM580 and TTNPB, histone methyltransferase (HMT) inhibitor EPZ004777, ascorbate, protein kinase C (PKC) inhibitor Go 6983, Rho-associated coiled-coil kinase (ROCK) inhibitor Y-27632, and c-Jun N-terminal kinase (JNK) inhibitor SP600125;
- wherein the fibroblasts are treated in the first composition for 5 days, and the fibroblasts are treated in the second composition for 7 days.
A method of preparing rejuvenated and regenerative fibroblasts, comprising:
- treating fibroblasts sequentially with a first composition and a second composition;
- wherein the first composition comprises HG-DMEM supplemented with 10% FBS and a first small molecule mixture consisting of ROCK inhibitor Y-27632, ascorbate, HMT inhibitor EPZ004777, cAMP agonist Forskolin and TGF-β inhibitor Repsox;
- and
- wherein the second composition comprises HG-DMEM supplemented with 10% FBS and a second small molecule mixture consisting of HDAC inhibitor VPA, WNT/β-catenin agonist CHIR99021, TGF-β inhibitor Repsox, cAMP agonist Forskolin, PKC inhibitor Go 6983, ROCK inhibitor Y-27632, RAR agonists AM580 and TTNPB, HMT inhibitor EPZ004777, and ascorbate;
- wherein the fibroblasts are treated in the first composition for 9 days, and the fibroblasts are treated in the second composition for 7 days.
Dependencies are inferred from the claim text.