University of Twente Student Theses
Hoffa's Fat Pad-on-Chip: Adipogenic differentiation of immortalized MSC's in Dextran - Hyaluronic acid - Gelatine hydrogels
Zandstra, T.J. (2024) Hoffa's Fat Pad-on-Chip: Adipogenic differentiation of immortalized MSC's in Dextran - Hyaluronic acid - Gelatine hydrogels.
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| Abstract: | Hoffa’s fat pad (HFP) is one of the articular fat pads in the knee joint. It has recently been shown that the HFP can play a role in the development of knee osteoarthritis. An accurate model of the HFP is therefore desired to study the cause of knee osteoarthritis. The Developmental Bioengineering group is currently developing a Hoffa’s fat pad-on chip combined with hydrogels. HFP mainly consists of adipocytes, which makes in vitro adipogenic differentiation in the hydrogel on chips important. This bachelor assignment will therefore focus on the ability of immortalized mesenchymal stem cells (MSC-hTERT) to undergo adipogenic differentiation in a 3D hydrogel environment. Firstly, the adipogenic differentiation ability of MSC-hTERT cells was tested in 2D culture. Regular adipogenic differentiation medium (A) was compared with higher insulin medium (B). Adipogenic stainings were performed to analyze the results. Oil Red O showed differention percentages of 46% and 41% for medium A and B respectively. Nile Red showed differentiation percentages of 66% and 68% respectively. Since the results were similar for both media, the medium with higher insulin was chosen as suggested by the creators of the cells. Secondly, the storage moduli of three cell densities in hydrogel were tested: 1 million/mL, 5 million/mL and 10 million/mL. The results showed a decrease in storage modulus as a result of increasing cell density, with 1 million/mL mimicking the in vivo situation best with a value of 611.58 Pa compared to adipose tissue with 713.2 Pa. Then, the adipogenic differentiation ability was tested in the 3D culture in hydrogels using 96-wells plates and Cartilage-on-chip platforms. The same three cell densities were tested in a 2.5% hydrogel composed of hyaluronic acid, dextran and gelatin. Both differentiated and undifferentiated cells showed cell viability over 19 days of culture, indicating high biocompatibility. Adipogenesis ability was best in the 5 million/mL hydrogels, with a differentiation percentage of 88%, compared to 44% in 1 million/mL and 63% in 10 million/mL, making it the most suitable for further development of the HFPoC. The chips were broken and did not give any staining results. Finally, actuation was tracked in chips seeded with beads. This showed a decreasing trend in displacement for a higher cell density, the further from the actuation membrane. These experiments indicate that a cell density of 1 million/mL mimicks the storage modulus of adipose tissue best, while a density of 5 million/mL provides the most adipogenic differentiation. |
| Item Type: | Essay (Bachelor) |
| Faculty: | TNW: Science and Technology |
| Subject: | 01 general works |
| Programme: | Biomedical Technology BSc (56226) |
| Link to this item: | https://purl.utwente.nl/essays/98211 |
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