公开(公告)号：US20230235289A1

公开(公告)日：2023-07-27

申请号：US17918981

申请日：2021-03-30

Applicant: Paul Scherrer Institut

Inventor： G.V. Shivashankar ,  Bibhas Roy

IPC: C12N5/077

CPC classification number: C12N5/0656 ,  C12N2506/1307 ,  C12N2527/00 ,  C12N2533/52 ,  C12N2533/54 ,  C12N2535/10

Abstract: Over the course of an aging process fibroblasts lose contractility, leading to reduced connective tissue stiffness. A promising therapeutic avenue for functional rejuvenation of connective tissue is reprogrammed fibroblast replacements with a laterally confined growth of fibroblasts on micro-patterned substrates that induces stem cell-like spheroids. The partially reprogrammed spheroids are embedded in collagen-I matrices of varying densities, mimicking different 3D tissue constraints. The spheroids regain their fibroblastic properties and sprout to form 3D connective tissue networks. The differentiated fibroblasts exhibit reduced DNA damage, enhanced cytoskeletal gene expression and acto-myosin contractility. The rejuvenated fibroblasts show increased matrix protein (fibronectin and laminin) deposition and collagen remodeling compared to the parental fibroblast tissue network. The partially reprogrammed cells have comparatively open chromatin compaction states and may be more poised to redifferentiation into contractile fibroblasts in 3D-collagen matrix. Collectively, the results highlight efficient fibroblast rejuvenation, with important implications in regenerative medicine.