NOVEL METHOD FOR HEMATOPOIETIC STEM CELL ISOLATION

这份小型企业创新研究第一阶段提案涉及 造血干细胞的纯化和分析问题。 目前用于干细胞分离的方法刺激增殖 具有细胞因子Kit配体（KL）的CD 34+骨髓细胞和 白细胞介素-3（IL-3）在5-氟尿嘧啶（一种细胞毒性药物）的存在下， 剂 通过这种处理使脱落细胞脱落， 基本上是纯的干细胞群体。 然而，很可能， 5-氟尿嘧啶对干细胞有一定的细胞毒性或致突变作用 如果要使用蒸汽室， 疗法 我们建议将CD 34+骨髓细胞包裹在 琼脂糖微滴，用KL和IL-3刺激增殖细胞， 并通过荧光染色和细胞分选纯化静止细胞。 分离的细胞将使用长期培养进行分析， 产生混合骨髓细胞的能力的细胞测定 血统 多能干细胞的研究具有新的紧迫性， 细胞工程的最新进展 开发新方法 选择和培养蒸汽细胞， 用于治疗和商业研究的技术。

This Small Business Innovation Research Phase I proposal addresses the problem of hematopoietic stem cell purification and analysis. current methods for stem cell isolation stimulate proliferative CD34+bone marrow cells with the cytokines Kit Ligand (KL) and interleukin-3 (IL-3) in the presence of 5-fluorouracil, a cytotoxic agent. Repliating cells are killled by this treatment leaving an essentially pure population of stem cells. However, it is likely that 5-fluorouracil has some cytotoxic or mutagenic effect on stem cell populations posing distinct problem if steam cells are to be used for therapy. We propose to encapsulate CD34+ bone marrow cells in agarose micro drops, stimulate proliferative cells with KL and IL-3, and purify quiescent cells by florescent staining and cell sorting. Isolated cells will be analyzed using a long term culture initiating cell assay for their ability to give rise to cells of mixed myeloid lineages. Research on pluripotent stem cells have taken on new urgency with recent advances in cell engineering. Development of new methods to select and culture steam cells is expected to advance the technology for therapeutic and commercial research use.