Presentation Title

Effects of Varying Adipose-Derived Stem Cell Concentrations on Chondrocyte Proliferation and Chondrogenesis in Co-Culture

Faculty Mentor

Mary Ziegler

Start Date

17-11-2018 12:30 PM

End Date

17-11-2018 2:30 PM

Location

HARBESON 18

Session

POSTER 2

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Achieving sufficient chondrogenesis from adult chondrocytes remains a prominent ongoing issue in tissue engineering. Human adipose-derived stem cells (hADSCs) promote chondrocyte proliferation and thereby possibly increase chondrogenesis. Thus, current avenues of research involve co-culturing chondrocytes with ADSCs. ADSCs may act upon chondrocytes via secreted factors, exosomes, and mitochondrial transfer and therefore interact with chondrocytes through both direct and indirect contact. Although, the relative importance of each form of contact is unknown. Additionally, the ratio of the combination of the two cell types in co-culture is not optimized. To explore the relationship between the two cell types in a co-culture environment and to optimize the cell ratios, we conducted both a transwell co-culture assay and a direct co-culture of hADSCs and porcine chondrocytes at three different ratios, with increasing concentrations of hADSCs: 1:3, 1:5, and 1:9 (chondrocytes:hADSCs). A CFSE cell proliferation assay was utilized to assess changes in chondrocyte proliferation, and the data were analyzed by flow cytometry with FlowJo software. Furthermore, to compare the effect of the varying ratios on chondrogenesis, tissue constructs seeded with the aforementioned ratios of cells were sectioned and stained with Alcian blue and were quantified with ImageJ software. The indirect co-culture of hADSCs and chondrocytes resulted in elevated proliferation compared to the direct co-culture. Altering the cell ratios did not affect proliferation; however, the 1:9 co-culture demonstrated the highest level of chondrogenesis, as shown by the Alcian blue staining results. These results indicated that direct contact between hADSCs and chondrocytes may simultaneously decrease proliferation and increase chondrogenesis. This work improves our understanding of the interaction between hADSCs and chondrocytes, and further analyses and optimization of the co-culture conditions will provide modes for improving cartilage yields, which is essential for expanding the current options for reconstructive surgeries requiring autologous cartilage.

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Nov 17th, 12:30 PM Nov 17th, 2:30 PM

Effects of Varying Adipose-Derived Stem Cell Concentrations on Chondrocyte Proliferation and Chondrogenesis in Co-Culture

HARBESON 18

Achieving sufficient chondrogenesis from adult chondrocytes remains a prominent ongoing issue in tissue engineering. Human adipose-derived stem cells (hADSCs) promote chondrocyte proliferation and thereby possibly increase chondrogenesis. Thus, current avenues of research involve co-culturing chondrocytes with ADSCs. ADSCs may act upon chondrocytes via secreted factors, exosomes, and mitochondrial transfer and therefore interact with chondrocytes through both direct and indirect contact. Although, the relative importance of each form of contact is unknown. Additionally, the ratio of the combination of the two cell types in co-culture is not optimized. To explore the relationship between the two cell types in a co-culture environment and to optimize the cell ratios, we conducted both a transwell co-culture assay and a direct co-culture of hADSCs and porcine chondrocytes at three different ratios, with increasing concentrations of hADSCs: 1:3, 1:5, and 1:9 (chondrocytes:hADSCs). A CFSE cell proliferation assay was utilized to assess changes in chondrocyte proliferation, and the data were analyzed by flow cytometry with FlowJo software. Furthermore, to compare the effect of the varying ratios on chondrogenesis, tissue constructs seeded with the aforementioned ratios of cells were sectioned and stained with Alcian blue and were quantified with ImageJ software. The indirect co-culture of hADSCs and chondrocytes resulted in elevated proliferation compared to the direct co-culture. Altering the cell ratios did not affect proliferation; however, the 1:9 co-culture demonstrated the highest level of chondrogenesis, as shown by the Alcian blue staining results. These results indicated that direct contact between hADSCs and chondrocytes may simultaneously decrease proliferation and increase chondrogenesis. This work improves our understanding of the interaction between hADSCs and chondrocytes, and further analyses and optimization of the co-culture conditions will provide modes for improving cartilage yields, which is essential for expanding the current options for reconstructive surgeries requiring autologous cartilage.