Development Of Continuous Density Gradient Cell Separati

开发了一种新的离心机系统，用于根据细胞密度连续分离细胞。它使用一个离心机碗，它在外围配备了一个圆形通道(3厘米宽，5毫米深，容量约为160毫升)，由隔膜中断，以便从一个终端引入的液体从另一端定量收集。该通道配备了6个进水口和6个出水口，每个出水口都连接到流管，这些流管使用无旋转密封装置通向离心机系统的外部。一组不同密度的聚合物悬浮液以1毫升/分钟的速度通过入口2至6引入，并以相同的流速通过各自的出口收集。样品细胞悬浮液以2-5ml/min的较高流速连续地通过第一通道进料，并通过相应的通风口收集。在离心力的作用下，样品悬浮液中的细胞逐渐迁移到聚合物层中，最终悬浮在相同密度的聚合物层中，然后到达通道的出口。人血黄大衣的初步结果表明，淋巴细胞和粒细胞溶解良好，红细胞与白细胞完全分离。CD34细胞主要分布在淋巴细胞部分(密度为1.060-1070)，这表明通过将树突梯度聚焦在1.065左右可以使它们变得更加丰富。处理1,000,000,000个细胞所需的时间约为3小时。最近，我们制作了一个类似但容量小得多的色谱柱，总容量只有8毫升。该系统可以用于少量细胞的分离，而聚合物悬浮液的成本要低得多。

A new centrifuge system is developed for continuous cell separation according to their densities. It uses a centrifuge bowl equipped with a circular channel (3 cm wide and 5 mm deep with an about 160 ml capacity) around the periphery which is interrupted by a septum so that the liquid introduced from one terminal is quantitatively collected from the other end. The channel is equipped with 6 inlets and 6 outlets each connected to flow tubes which are led to the outside of the centrifuge system using a rotary-seal-free device. A set of polymer suspensions of different densities is introduced through inlets 2 to 6 at 1 ml/min and collected through the respective outlets at the same flow rates. The sample cell suspension is continuously fed through the first channel at a higher flow rate of 2 - 5 ml/min and collected through the respective outlet which is open to the air. Being subjected to a centrifugal force cells in the sample suspention gradually migrate into the polymer layers and are finally suspended in the polymer layer of the same density before reaching the outlet of the channel. The preliminary results obtained from human blood buffy coat revealed that the lymphocyts and granulocytes are well resolved and erythrocytes are completely separated from white cells. CD34 cells are mainly distributed in the lymphocyte fraction (density 1.060-1070) suggesting that they may be enriched by finely focusing the dendity gradient around 1.065. The time required to process 1,000,000,000 cells are about 3 hours. More recently we have made a similar but much smaller capacity column with only a 8 ml total capacity. The system can be used for the separation of a small number of cells with much less cost for the polymer suspension.