Syva Hednella Sabanting (2017) Cellular interaction between human amnion Mesenchymal stem cells, human umbilical vein endothelial cells and human dermal fibroblasts in 2D and 3D in vitro triculture systems. Masters thesis, Universiti Malaysia Sabah.
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Abstract
Human amnion mesenchymal stem cells (HAMCs) are capable of multilineage differentiation and have angiogenic potential. Tri-culture system is known to direct stem cell fate decisions. This study was carried out to determine the cellular interaction between HAMCs, human umbilical vein endothelial cells (HUVECs) and human dermal fibroblasts (HDFs) in 2-dimensional (2D) and 3-dimensional (3D) tri-culture systems in vitro. In the first part of the study, HAMCs, HUVECs and HDFs were cultured directly and indirectly in 2D tri-culture system at various seeding ratio (HAMCs:HUVECs:HDFs; 1:4.5:4.5, 5:2.5:2.5 and 9:0.5:0.5) until 7 days. Cell morphological observation, ELISA for vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) detection and immunofluorescence staining against von willebrand factor (vWF), stage specific embryonic antigen-4 (SSEA-4) and collagen Type I ( coll) were performed. In the second part of the study, HAMCs, HUVECs and HDFs were embedded in fibrin scaffold at selected seeding ratio, 9:0.5:0.5 for 7 days. Cells morphological observation, ELISA and immunohistochemistry against SSEA-4, monoclonal mouse anti-human CD31 and alpha smooth muscle actin (SMA-a) were performed. The results in 2D study showed that HAMCs retained its spindle-shaped morphology with filopodia, and VEGF concentration in both direct and indirect tri-culture groups was significantly reduced at the end of culture whereas bFGF concentration remained constant. HAMCs in both direct and indirect 2D tri-culture positively expressed against SSEA-4 and Coll but negatively expressed vWF. Whereas in 3D study, HAMCs in tri-culture formed network- and capillary-like structures and showed significant reduction in VEGF and bFGF concentration on Day 7. They positively expressed SMA-a but negatively expressed SSEA-4 and CD31. The results in 2D study suggested that HAMCs do not differentiate into endothelial cell. However, VEGF molecules was activated to promote proliferation and migration. For 3D tri-culture study, HAMCs morphology demonstrated its involvement in angiogenesis, and differentiated into myofibroblasts cells. HAMCs in 2D tri-culture maintain its MSCs characteristic but not endothelial cell whereas HAMCs in 3D tri-culture do not possessed endothelial cell and MSCs characteristics. HAMCs in 2D do not influence and be influenced by other cell in direct and indirect contact. However, HAMCs differentiated and proliferated in the presence of fibrin. Therefore, the features of 3D tri-culture system model in this study have expanded the limitation of 2D tri-culture system. The 3D system has successfully provided fundamental knowledge of mimicking skin microenvironment for future work as well as future application towards in viva study. Nevertheless, further fundamental studies are required to assess HAMCs differentiation and roles in angiogenesis in 2D and 3D in vitro tri-culture models.
| Item Type: | Thesis (Masters) |
|---|---|
| Keyword: | Human amnion mesenchymal stem cells, HAMCs, Tri-culture system, Angiogenesis, Endothelial cells, 3D culture, Myofibroblasts, VEGF, Fibrin scaffold |
| Subjects: | Q Science > QH Natural history > QH301-705.5 Biology (General) > QH573-671 Cytology |
| Department: | INSTITUTE > Biotechnology Research Institute (BRI) |
| Depositing User: | DG MASNIAH AHMAD - |
| Date Deposited: | 11 Jun 2025 16:04 |
| Last Modified: | 11 Jun 2025 16:04 |
| URI: | https://eprints.ums.edu.my/id/eprint/44115 |
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