Application of relatively hydrophobic strategies and approaches to construction of functional tissue constructs
Date Issued
2010
Date
2010
Author(s)
Hsieh, Chin-Hsiung
Abstract
Relative hydrophobic culture strategy and robotic laser micromachining were used in this study to develop a simple and automatic process of cell array. This method has been applied in the research of cell-cell interaction and the response of cell to drug treatment. Furthermore, the relative hydrophobic culture strategy and the microwell cell patterning provided a suitable environment for cultivating self-assembled spheroidal dermal papilla microtissues in the commercial polystyrene culture plate. The arrayed functional microtissues of dermal papilla can be applied to the reconstruction of hair for patients with alopecia.
In the first experiment, we designed a strategy to improve the efficiency of cell adhesion to the scaffold using a hydrophobic cell culture environment Cells show lower affinity to the surface of PDMS than tissue culture polystyrene (TCPS) plate. When cells were cultured with gelatin microspheres or chitosan films in PDMS-coated plate instead of a normal TCPS plate, there was a significant increase in cell attachment efficiency. The results demonstrate that the method is easy to use and facilitates fast cultivation of cell-scaffold constructs.
In the second experiment, we developed a method for fast cell array fabrication using laser sintering and the hydrophobicity of PDMS films (Patterned PDMS-based cell array system, PCAS). This approach can be easily adopted and is cost-effective. We used NIH/3T3 fibroblast cells to demonstrate the feasibility of PCAS. We also used PC-12 cells with poor adhesion ability to demonstrate cell-cell communication. Results showed that the method is very useful for studying cell-cell interaction, cell-substrate interaction or cell migration.
Finally, we employed the PCAS to develop a strategy for cultivating dermal papilla (DP) cells to form multiple arrayed spheroidal microtissues for transplantation. By controlling the cell seeding densities, a microwell with arrayed DP spheroidal microtissues was easily formed. Formation of DP microtissues was associated with the overlapping of multilayered cells on microwells and low cell-substrate adhesivity on the PDMS film. A microwell environment enhanced the aggregation of DP cells into spheroidal microtissues on the TCPS culture plate. The aggregation of DP microtissues formed on microwells preserves their hair induction potential for use in follicular cell implantation and is independent of the number of cells in the TCPS plate. Large quantity of DP spheroidal microtissues can be obtained fast and simply by the platform.
Subjects
PDMS
tissue engineering
cell array
dermal papilla
hair follicles
Type
thesis
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