Study on Fluorescence Activated Micro Cell Sorter with On-Chip Cell Expansion System
Date Issued
2007
Date
2007
Author(s)
Wang, Yen-Siang
DOI
zh-TW
Abstract
A new microfabricated platform for rare cells is demonstrated, which provides specific cells with a novel device of multiple functions of cell sorting, cell culturing, and real-time investigation. Due to increasing potential of stem cell in therapy, diagnostics, drug discovery as well as development of its science and technology, few existing techniques can be tailored to be specific for emerging needs of stem cell applications. In primary culture of rare stem cells that account for one out of 103~106 cells, purification and expansion of stem cells requires complicated processes of cell identification, sorting, culture and verification. The present microfabricated platform that provides integration of required functions is especially suitable for purification of stem cells in mixed population of rare cells. The micro device is designed to be disposable for single use and minimal contamination for potential cell therapy. In addition, the single-use device is able to be compatible with the existing microscopes and to be capable of integration with subsequently required cell analysis.
The micro device is simply composed of a polydimethylsiloxane (PDMS) elastomer channel and multiple electrodes deposited on the glass substrate. By using PDMS, a controllable environment for target cells with bio-compatible (hydrophobic), elastic vessel-like channel wall, and transparent (optically transparent down to 300 nm) for microscopic living-cell detection was created. In sorting approach, the dielectrophoresis (DEP) technique with hydrodynamic focusing flow is chosen due to minimal effect to cells and ease of implementation. The moving fluorescent cells tracked under the microscope to be sorted were guided along the designated electrodes by DEP to the specific chambers, while the rest of cells were straight transported through the central flow of the hydrodynamic focusing to the waste chamber. Few amount of the selected cells were required for proliferation on chip to be contamination-free. To facilitate the on-device culture of living cells, the culture condition was additionally controlled by a micro-incubator controller, providing an excellent controlled environment at just 37℃, 90% moisture and 5% CO2 concentration. Several types of human cells chosen for preliminary studies enable the present device to simulate direct applications on clinical trial.
The present system demonstrates the potential for surface marker-specific cell sorting of human bone marrow stem cells simply with the normal, regular procedures of immunostaining techniques by using μ-FACS within the micro device. To evaluate the performance of this on-chip culture system shown, the sorted human bone marrow stem cells had been examined for detailed testing. The cell-sorting performance of rare cells shows 92 % in efficiency. Over one day and several hours of culture, the sorted cells exhibited a well-adhered, healthy morphology comparable to static culture and the surface of untreated glass was almost completely covered by the proliferating cells.
The results demonstrated that the microchamber is a satisfactorily biocompatible microenvironment for the continuous growth of the cells. The multifunctional μ-platform system has been proven to be useful in rare stem cells sorting and cell culturing. The platform offers the potential for the future application on stem cell therapy.
Subjects
細胞分選
介電泳
水力聚焦
罕見細胞
幹細胞
cell sorting
dielectrophoresis
DEP
hydrodynamic focusing
rare cell
stem cell
SDGs
Type
thesis
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