Mechanical and molecular parameters that influence the tendon differentiation potential of C3H10T1/2 cells in 2D- and 3D-culture systems

Abstract

One of the main challenges relating to tendons is to understand the regulators of the tendon differentiation program. The optimum culture conditions that favor tendon cell differentiation have not been identified. Mesenchymal stem cells present the ability to differentiate into multiple lineages in cultures under different cues ranging from chemical treatment to physical constraints. We analyzed the tendon differentiation potential of C3H10T1/2 cells, a murine cell line of mesenchymal stem cells, upon different 2D-and 3D-culture conditions. We observed that C3H10T1/2 cells cultured in 2D conditions on silicone substrate were more prone to tendon differentiation, assessed with the expression of the tendon markers Scx, Col1a1 and Tnmd as compared to cells cultured on plastic substrate. The 3D-fibrin environment was more favorable for Scx and Col1a1 expression compared to 2D cultures. We also identified TGFβ2 as a negative regulator of Tnmd expression in C3H10T1/2 cells in 2D and 3D cultures. Altogether, our results provide us with a better understanding of the culture conditions that promote tendon gene expression and identify mechanical and molecular parameters upon which we could act to define the optimum culture conditions that favor tenogenic differentiation in mesenchymal stem cells.

Core–Shell Pure Collagen Threads Extruded from Highly Concentrated Solutions Promote Colonization and Differentiation of C3H10T1/2 Cells

“…Noteworthy, in situ hybridization also reveals that whether in the sheath around or penetrated in the threads, cells show a strong differentiation. The profile of differentiation is similar to that of C3H10T1/2 cells cultivated within fibrin hydrogels except for the collagen expression 45 . Hence, collagen threads allow for a sustainable collagen I expression by cells.…”

“…The profile of differentiation is similar to that of C3H10T1/2 cells cultivated within fibrin hydrogels except for the collagen expression. 45 Hence, collagen threads allow for a sustainable collagen I expression by cells. As a result, these cells adopt a biosynthetic profile required for tendon regeneration.…”

Dynamic Load Model Systems of Tendon Inflammation and Mechanobiology

“…(B) Tenocyte cell culture monolayers (2D in vitro systems) under static load, or no load, and dynamic loads have the greatest high-throughput capabilities. Dynamic loads on flexible membranes may be produced by mechanical actuators or with vacuum pressure, as illustrated and modified from Flexcell International's Tension System, Burlington, NC (Arnoczky et al, 2002;Fleischhacker et al, 2020;Gaut et al, 2020;Kubo et al, 2020). (C) Cell-seeded 3D constructs can be gel-like, as shown in the side view (modified from Flexcell's 3D Tissue Train System), or like whole-tissue if decellularized constructs are used.…”

A novel fabrication process of up‐scalable microfiber‐shaped tendon‐like tissue with high cell density for uniformed macroscale assembly

“…The relative gene expressions of tenomodulin (Tnmd), a marker of tendon differentiation that promotes tendon proliferation and collagen fibril maturation (Gaut et al, 2020), Mohawk (Mkx), a gene to promote regeneration of tendon (Milet & Duprez, 2015), and Scleraxis (Scx) a marker that regulates the differentiation of tendon (Blitz et al, 2009), were determined and normalized by ribosomal protein S18 (Rps18), a housekeeping gene. The gene expressions of Day 14…”