【作者】 韩林峰；

【导师】 沈鹤柏；

【作者基本信息】 上海师范大学 ， 物理化学， 2009， 硕士

【摘要】 造血干细胞自动磁分选仪采用微型计算机和现代控制技术,通过磁性纳米粒子与CD34单克隆抗体结合,构建磁性纳米粒子生物分子复合物,制成细胞磁分选溶液,进行造血干细胞分离。对分离过程和管道清洗进行全程自动控制,可以批量、安全、精确、快速地对干细胞进行分离,用于从外周血或脐带血中分离出造血干细胞。采用该分离系统后,可以使造血干细胞分离成本成倍地降低。用于临床后可为众多的肿瘤和免疫疾病的患者带来治疗和治愈的机会,用该分选系统分选后,在异体PBSCT中它可去除T细胞、减少和减轻GVHD发生,这种大量细胞磁分选仪采用微型计算机和现代控制技术,对细胞分离和洗涤过程进行全程自动控制,可以批量、安全、高效,快速地对干细胞进行分离。本论文主要取得了以下结果:(1)造血干细胞自动磁分选仪采用微型计算机和现代控制技术,对细胞分离和洗涤过程进行全程自动控制。根据仪器的功能及应用,设计该仪器的硬件系统包括磁分选系统,样品传送系统、电脑控制系统和外壳。硬件设计主要完成了分选柱研制、管道设计、外壳设计等工作。造血干细胞自动磁分选仪的硬件系统设计全面、实用,作为一台医疗研究仪器除了具有性能优越,精确耐用等硬性指标外,更具备美观大方,便于操作的特色。(2)造血干细胞自动磁分选仪是通过电脑编程实现全自动化控制的,电脑控制系统可分为:电磁阀控制、磁块电机控制、蠕动泵控制,每个控制系统又细化为几个部分。控制系统软件编程时要求对仪器运行的每一步骤有精确的描述和表达。仪器运行分为两个大的步骤,一为系统测试和管道润洗步骤,该步骤有34步命令,历时2分钟;一为细胞自动分选步骤,该步骤有89步命令,历时20分钟。整个仪器运行状态时间为25分钟左右。造血干细胞自动磁分选仪的分选从冲洗系统前滤器开始,然后开始标记细胞的分选。(3)造血干细胞自动磁分选仪采用微型计算机和现代控制技术。实验通过磁性纳米粒子与CD34单克隆抗体结合,构建磁性纳米粒子—生物分子复合物,制成细胞磁分选溶液。磁性纳米粒子大部分粒径都在50-80nm,微粒的分布窄,是混合型包裹,里面包裹了好几个磁性纳米粒子,分散性也比较好,团聚现象不是很严重,具有良好的超顺磁性。在造血干细胞自动磁分选仪上进行造血干细胞分离富集,得到了较好的试验结果,经过流式细胞仪检测获得分离纯度50%以上,分离效率40%以上,体外扩增效果良好,具备较强的实用性。更多还原

【Abstract】 Hematopoietic stem cell automatic magnetic separation instrument, which we design and development, uses microcomputer and modern control technology. Through a magnetic nanometer granule and the CD34+ monoclonal antibody union to construct the magnetic nanoparticles biological molecular complexes, it makes the magnetic cell sorting solution for hematopoietic stem cell separation. Carrying on the full automatic control to the separation process and the pipe clean, may separate the stem cell batch, safely, precisely and fast. in order to isolate stem cells for the blood or cord blood from the isolated hematopoietic stem cells. Using this separation system, may cause to make the cost of blood stem cell separation reduce multiplied. Can be used in clinical as many tumors and autoimmune diseases in patients brought treatable and curable, use of the election system after the election, in which it allogeneic T cells can be removed to reduce GVHD occurred. Such large magnetic cell sorting adopts micro computer control and modern technology, on the cell separation and washing entire process control, batch, safe, efficient and rapid method to isolate cells.This paper mainly includes two aspects as follows:(1) Hematopoietic Stem Cell Sorter automatically using magnetic micro-computersand modern control technology, cell separation and washing the whole process automatic. According to the function and application of equipment, design of the hardware system of the apparatus including magnetic separation system, the sample delivery systems, computer control system and shell. Hardware Design of the main separation column completed development, pipeline design, casing design and so on. Hematopoietic Stem Cell Sorter automatically magnetic hardware system design a comprehensive, practical, as a medical research equipment In addition to superior performance, durability and other hard targets accurately, but also have the elegant, user-friendly features.(2) Hematopoietic Stem Cell Sorter automatically magnetic computer programmingthrough the control of fully automated, computer-controlled system can be divided into: control solenoid valve, magnetic motor control block, control of peristaltic pump, each control system for parts and refinement. Control system software to run when the requested equipment for each step and describe the precise expression. Instrument operation is divided into two major steps, one for system testing and pipe run washing steps, the step command steps to 34, spend 2 minutes. Second, cellular automata for sorting steps, the step command steps to 89, spent 20 minutes. Machines running the entire time is 25 minutes. Hematopoietic Stem Cell Sorter automatic magnetic separation wash system from the beginning of pre-filter, and then began sorting the labeled cells.(3) Hematopoietic Stem Cell Sorter automatically using magnetic micro-computers and modern control technology. Experiment through the magnetic nanoparticles with CD34 monoclonal antibody with magnetic nanoparticles to build - complex biological molecules, made of magnetic cell sorting solution. Most of magnetic nanoparticles in the size 50-80nm, narrow distribution of particles is a hybrid package, which packages a number of magnetic nanoparticles, dispersion also better reunion phenomenon is not very serious, a good super-paramagnetic. Hematopoietic stem cells in the automatic magnetic separation apparatus for separation and enrichment of hematopoietic stem cells, have been a better test results, after the separation by flow cytometry to obtain more than 50%purity, more than 40% separation efficiency, good effect in vitro amplification with more practical.更多还原