Establishment of fibroblast and myofibroblast phenotypes for use in in vitro co-culture models.

Autor: Ramklowan DSH; Discipline of Biochemistry, School of Life Sciences, University of KwaZulu Natal, Scottsville, South Africa., Snyman C; Discipline of Biochemistry, School of Life Sciences, University of KwaZulu Natal, Scottsville, South Africa., van de Vyver M; Division of Clinical Pharmacology, Department of Medicine, Faculty of Medicine and Health Sciences, Stellenbosch University, South Africa., Niesler CU; Discipline of Biochemistry, School of Life Sciences, University of KwaZulu Natal, Scottsville, South Africa. Electronic address: niesler@ukzn.ac.za.
Jazyk: angličtina
Zdroj: Biochimie [Biochimie] 2023 Apr; Vol. 207, pp. 96-101. Date of Electronic Publication: 2022 Nov 04.
DOI: 10.1016/j.biochi.2022.10.017
Abstrakt: Fibroblasts function to secrete and modify components of the extracellular matrix. During wound healing, fibroblasts migrate to the site of injury and differentiate into contractile myofibroblasts; this differentiation is characterised by an increased contractile capacity. Fully differentiated myofibroblasts can be distinguished from fibroblasts via the higher expression of α-smooth muscle actin as well as a denser cytoskeleton. Impaired wound healing has been characterised by a lack of myofibroblasts; as a result, tissue does not fully regain its strength and function. Under pathological conditions, this may be associated with the effect that a pro-inflammatory microenvironment has on fibroblast and skeletal muscle progenitor cell migration and differentiation. Given their distinct roles in tissue maintenance and repair, the communication between fibroblasts versus myofibroblasts with other cellular mediators of repair is likely to influence cell behaviour and the outcome of wound repair. An in vitro test model is required to investigate this intercellular influence, but the establishment of such a model is hampered by the difficulty in retaining the dedifferentiated fibroblastic phenotype under regular serum-containing cell culture conditions. We present a model that supports the establishment and retention in culture of fibroblast and myofibroblast phenotypes for use in a simple, inexpensive, yet relevant in vitro 2D assay. This model is then applied in a co-culture setting to determine whether the presence of myoblasts affects the ability of fibroblasts versus myofibroblasts to close an in vitro wound. Our results emphasize the importance of considering the impact of paracrine communication between all cells during wound healing.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2022 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)
Databáze: MEDLINE
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