Glioma Stem Cells 3D Culture And The Exploration Of Aptamers Selection

Objective: 1. The three-dimensional(3D) cell culture techniques are adopted to simulate the environment heterogeneity in vivo to study glioma stem cells proliferation and some biological characteristics. 2. Synthetic DNA aptamer library synthesis in vitro, using Cell- SELEX technology screening, screening can get specific identification of nucleic acid aptamer glioma stem cells.Methods: 1.Using microporous micro array chip technology to produce high flux collagen array, and then press the way of using a fixed force to make the cells were distributed evenly in the array, form a three-dimensional culture model. 3D cultivation conditions for a long time, high flux glioma stem cell heterogeneity analysis. 2. Using random oligonucleotide library of chemical synthetic method, the cultured human glioblastoma cell line U251 cells of glioma stem cells as target cells in vitro, human glioblastoma cell line U251 cell as the against cell, using the Cell- SELEX technology, screen DNA aptamer.Results: 1, The glioma cells can be collagen high-throughput microarray array for medium and long-term survival, proliferation, and cell activity has no effect. 2, After 7 days, 30 days cultured, observation and evaluation, 3 D culture under the condition of glioma stem cells and glioma cells can be observed between the proliferation of differences and heterogeneity in the process of each cell to extend. 3, After 15 rounds of selex,nucleic acid aptamer library can combine glioma stem cells 4, The nucleic acid aptamer libraries can well combine with the human brain glioma stem cells, but not with the brain glioma cell line U251 combination.Conclusion:1, We establish a high-throughput single-cell 3D cultivation platform, analysis for glioma stem cell heterogeneity. Glial tumors cells extend has limitations and the glioma stem cells can almost infinite proliferation. 2, The library that Screened can be combined with specificity in nucleic acid adaptation target protein molecules on the surface of the target cells, to fit the body in the search for biomarkers of glioma stem cells, as well as gliomas targeted drug, postoperative chemotherapy, intraoperative navigation research laid a solid foundation in the field of application.