Release date: 2014-08-05

Researchers from the National Institutes of Health (NIH) have recently developed a technology that uses this technology to accelerate stem cell formation, a technology that simultaneously measures the expression of multiple genes, helping researchers to cell based on cell function and developmental stages. classification. For example, the use of this technology will help researchers use the patient's skin cells to regenerate retinal pigment epithelial cells (RPE, tissue behind the eyeball, many blind areas that are damaged), which will also help researchers find personalized medications. The findings were published in the recent issue of Stem Cell Translational Medicine.

RPE is a single-cell layer located near the retina, which is covered with cones and rods containing photoreceptors. RPE is critical for sensitization, and many diseases that cause RPE damage can cause vision and sensitization. Loss.

Artificially induced pluripotent stem cell technology (iPS) promotes the formation of RPE by stem cells. To validate the RPE produced by iPS cells, the researchers used microscopy and physiological function tests to ensure that the morphology and function of the regenerated tissue were normal. The researchers also used quantitative RT-PCR to detect gene expression levels that reflect cell developmental processes and functions. For example, in iPS cells, SOX2 gene expression is higher than mature RPE, but quantitative RT-PCR only allows researchers to Each sample was tested simultaneously for several genes. The research team developed a new multi-measurement chip based on the commercial platform of biotechnology company Affymetrix. The chip is highly automated to measure the expression of multiple genes on RPE samples. In the chip, small DNA fragments are bound to the beads to capture the RNA molecules expressed by the genes of the cells in the RPE sample. Once acquired, RNA molecules from different genes are labeled with fluorescent labels.

The researchers used iPS technology to induce RPE generated by skin tissue, and then detected the expression of eight marker genes representing development, function, and disease in RPE. The team used quantitative RT-PCR to simultaneously detect RNA levels in each gene, then simultaneously detected all genes using a multi-channel chip and compared gene expression differences between RPE and iPS cells and RPE cells produced by iPS. Compared to iPS cells, iPS produced lower levels of SOX2 in RPE, while RPE-specific genes PAX6, RPE65, RDH5, TRPM1 and BEST1 genes were higher. Compared to the cultivated RPE, the iPS produced RPE with higher levels of SOX2 and PAX6 gene expression (the two genes are development-related markers), but the RPE-specific gene level is lower, suggesting an immature state.

The researchers say the work demonstrates that automated high-throughput technology can be used to simultaneously measure multiple gene expression profiles of RPE produced by iPS, which means savings, increased efficiency and yield. Multi-channel chips will be applied to cell quality checks and used to screen for potentially potent compounds and will also help improve RPE-related cell therapy.

Source: Bio-Exploration

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