【摘 要】
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Purpose:Mouse lung multipotent stem cells (MLSCs) could regenerate many different lung components including bronchioles, alveoli, and pulmonary vessels.But the research of MLSCs were limited due to it
【机 构】
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State Key Laboratory of Trauma,Bums and Combined Injury,Institute of Surgery Research,Daping Hospita
【出 处】
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首届全国组织修复与再生医学大会暨国际组织修复与再生医学学术研讨会
论文部分内容阅读
Purpose:Mouse lung multipotent stem cells (MLSCs) could regenerate many different lung components including bronchioles, alveoli, and pulmonary vessels.But the research of MLSCs were limited due to its rarity.In this study, we introduced a novel microfluidic magnetic activated cell sorting system in the isolation of MLSCs.Materials and Methods The CD45-pos and CD31-pos cells (hematopoietic cells and endothelial cells) are magnetically labled with anti-murine CD45 antibody-coated and anti-murine CD31 antibody-coated microbeads.Then, the single cell suspension is loaded onto the Sample Inlet of the cartridge which is then placed in the sample tray of IsoFluxTM.The sample flows into isolation zone at low velocities and sees an externally applied magnetic field from above the collection substrate.CD45-pos and CD31-pos cells with magnetic beads are attracted to the roof of the isolation zone, whereas other unlabled cells are biased from flow and gravitational forces to continue down the channel and into the Unlabled Cell Zone.CD45-neg and CD31-neg cells were then incubated with anti-mouse Sca-1 antibody conjugated to FITC, after which magnetic labeling of Sca-1-pos cells can be achieved using Anti-FITC MicroBeads.Then, cells were flowed through the channel at a flow rate of 20 μl per minute by applying a head pressure of 2 psi to the inlet well.Cells flow into isolation zone and Sca-1-pos cells with magnetic beads are attracted to the roof of the isolation zone.The roof of the isolation zone is a polymeric disk that is separate from the construction of the main flow channel.After processing, the disk containing the cells in a hanging drop is inserted into a tube.
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