Conditioned media cause increases in select osteogenic and adipogenic differentiation markers in mesenchymal stem cell cultures

Abstract

The optimal mesenchymal stem cell (MSC) culture conditions that would allow clinically viable tissue-engineered devices are still yet to be determined. Most MSCs are found in the bone marrow, an area that also contains numerous osteoblasts and adipocytes. Paracrine signalling may be leveraged to modulate MSC differentiation in the preparation of a tissue-engineered device. Thus, the objectives of this study were to determine the effects of adipocyte-conditioned medium (CM) on MSC differentiation to osteoblasts and to determine the effects of osteoblast CM on MSC differentiation to adipocytes. Two groups of murine MSCs were given either an osteogenic differentiation medium or an adipogenic differentiation medium. CM was taken from one group and administered to the opposite group in concentrations of 25% or 50%. Metabolic activity, total protein and alkaline phosphatase (ALP) assays were conducted on the osteogenic group at predefined time points throughout the 21 day study, while metabolic activity, triglyceride and oil red O assays were conducted on the adipogenic group at predefined time points. Adipocyte CM administered at a concentration of 50% increased the ALP production of MSCs undergoing osteogenic differentiation. Additionally, osteoblast CM increased the triglyceride production of MSCs undergoing adipogenic differentiation and enlarged the lipid vesicles that were produced by the cells. The effects of the osteoblast CM were seen at both concentrations, but were greatest at the 50% CM level.

Three-Dimensional In Vitro Tri-Culture Platform to Investigate Effects of Crosstalk Between Mesenchymal Stem Cells, Osteoblasts, and Adipocytes

“…Taken together, this suggests that co-cultured MSCs, while not terminally differentiated, may be biased toward differentiation toward the cell type with which they were cocultured, in agreement with previously published data from other groups. [22][23][24] Tri-cultured (OMA) MSCs, while closely related to osteoblasts and OMO MSCs in their gene expression and apparent alkaline phosphatase expression (Figs. 4A, B and 5A, B), correlated with a broader set of mesenchymal lineage genes, including MYOD (Fig.…”

“…16,21 Conditioned media and 2D co-culture studies have provided some insight into how one cell type (osteoblasts or adipocytes) affects the function and differentiation of MSCs. [22][23][24] However, these experiments only model static, one-way interactions and there is currently no systematic means to examine the effects of multidirectional and dynamic crosstalk over time between multiple cell types simultaneously in 3D culture in a way that better mimics interactions that occur in vivo.…”

Preparation and characterization of starch-poly-ε-caprolactone microparticles incorporating bioactive agents for drug delivery and tissue engineering applications

“…Dexamethasone (DEX) has been widely used in clinical applications to treat immuno-disorders [41,42], but a more specific and common use has been the control of the inflammatory response and tissue repair during organ transplantation [43]. In the last years, the use of this corticosteroid as an osteogenic agent has increased considerably in in vitro cell culture to induce the differentiation of stem cells into an osteoblastic lineage [41,[44][45][46].…”

Enhanced Proliferation and Accelerated Adipogenic Differentiation of Mesenchymal Stem Cells via Fe3O4-Insulin Conjugates Stimulation

“…Under appropriate microenvironments, MSCs can differentiate into multiple cell types, including adipocytes, chondrocytes, and cardiomyocytes, also into non-mesodermal-derived cells, including hepatocytes and neurons [3][4][5][6]. Recently, MSCs are being evaluated for their use in tissue engineering and cell-based therapies because of their no ethical restriction and low immunogenicity [7,8].…”