Effect of Cell Seeding Density and Inflammatory Cytokines on Adipose Tissue-Derived Stem Cells: an in Vitro Study

Abstract

Adipose tissue-derived stem cells (ASCs) are known to be able to promote repair of injured tissue via paracrine factors. However, the effect of cell density and inflammatory cytokines on the paracrine ability of ASCs remains largely unknown. To investigate these effects, ASCs were cultured in 8000 cells/cm2, 20,000 cells/cm2, 50,000 cells/cm2, and 400,000 cells/cm2 with and without 10 or 20 ng/ml tumor necrosis factor alpha (TNFα) and 25 or 50 ng/ml interferon gamma (IFNγ). ASC-sheets formed at 400,000 cells/cm2 after 48 h of culture. With increasing concentrations of TNFα and IFNγ, ASC-sheets with 400,000 cells/cm2 had increased production of angiogenic factors Vascular Endothelial Growth Factor and Fibroblast Growth Factor and decreased expression of pro-inflammatory genes TNFA and Prostaglandin Synthase 2 (PTGS2) compared to lower density ASCs. Moreover, the conditioned medium of ASC-sheets with 400,000 cells/cm2 stimulated with the low concentration of TNFα and IFNγ enhanced endothelial cell proliferation and fibroblast migration. These results suggest that a high cell density enhances ASC paracrine function might beneficial for wound repair, especially in pro-inflammatory conditions.Electronic supplementary materialThe online version of this article (doi:10.1007/s12015-017-9719-3) contains supplementary material, which is available to authorized users.

Nanofibrillar cellulose wound dressing supports the growth and characteristics of human mesenchymal stem/stromal cells without cell adhesion coatings

“…In contrast, we discovered unchanged gene expression levels of FGF2, TNF-α, and IL-6 in hASCs cultured on type 3 NFC dressing compared with control cells when using the higher cell density. Sukho et al have shown that cell seeding density affects greatly to the cytokine and growth factor secretion levels of hASCs [49]. Further, Fig.…”

“…Type 3 NFC dressing did not induce any remarkable cytotoxicity [46], and hASCs adhered to the surface of type 3 NFC dressing within 24 h and showed similar morphology compared with the control cells after 1-week culturing. However, the used cell density was excessively higher compared with the traditionally used cell densities that have been identified to affect the growth and functionality of cells [47][48][49]. In specific, high cell density has been shown to promote paracrine action of hASCs [49], which might benefit the wound repair process.…”

“…However, the used cell density was excessively higher compared with the traditionally used cell densities that have been identified to affect the growth and functionality of cells [47][48][49]. In specific, high cell density has been shown to promote paracrine action of hASCs [49], which might benefit the wound repair process. Nevertheless, hASCs maintained high cell viability during the 2-week culturing.…”

Rat Mesenchymal Stromal Cell Sheets Suppress Renal Fibrosis via Microvascular Protection

“…Therefore, in contrast to other studies, IV administration of BMSCs did not show any effects in terms of kidney function based on serum Cr and blood urea nitrogen. Since paracrine effects of MSCs depend on the number of cells , therapeutic effects are expected to be stronger when the number of transplanted cells increase. However, at the same time, mortality increases due to pulmonary embolism.…”

Patient-derived small intestinal myofibroblasts direct perfused, physiologically responsive capillary development in a microfluidic Gut-on-a-Chip Model

“…Second, because cell culture density is known to have a substantial effect on experimental outcome owing to paracrine signaling 28 , we tested the effect of altering cell concentrations of ISEMFs and ECs. We investigated four different loading cell densities, each sequential density being double to the one prior ( Fig.…”