A new continuous‐flow cell separation method based on cell density: Principle, apparatus, and preliminary application to separation of human buffy coat

Abstract

With the recent progress in transfusion medicine, separation and isolation of cells in a large quantity is becoming increasingly important. At present, the continuous cell separation method in a preparative scale is limited to apheresis and elutriation: the former is mainly used for collection of platelet and buffy coat from the whole blood, while the latter separates cells virtually according to their size. Here we introduce a continuous flow method that separates cells entirely based on cell density. The method is gentle and capable of processing a large number of cells. The potential capability of the method was demonstrated on separation of lymphocytes and granulocytes from human buffy coat. Lymphocytes were enriched to 90% in the fraction at density = 1.065 and granulocytes are isolated in fractions at density = 1.075-1.080 while red cells were completely retained at the periphery of the channel. CD34 cells were distributed around 1.065 and coeluted with lymphocytes, suggesting that further enrichment requires focusing the density gradient around 1.070. The method could process 10(9) nucleated cells in 2 hours. Our preliminary results suggest that the present method is an effective and efficient means to separate blood cells.

Synthetic Nano- and Micromachines in Analytical Chemistry: Sensing, Migration, Capture, Delivery, and Separation

“…In addition, these techniques separate species based on differences in their physical properties such as shape (150), density (151), or charge (152). This presents a problem in multicomponent sorting, given the physical difference between particles is often not enough for efficient separation (151). This problem may be overcome by separating species on the basis of their self-powered propulsion and chemotaxis.…”

“…Most current separation techniques at the nanoor microscale use continuous flow devices, which often require an external force field. In addition, these techniques separate species based on differences in their physical properties such as shape (150), density (151), or charge (152). This presents a problem in multicomponent sorting, given the physical difference between particles is often not enough for efficient separation (151).…”

Label-free, microfluidic separation and enrichment of human breast cancer cells by adhesion difference

“…7 These cell sorting systems have been miniaturized to microfluidic devices, demonstrating the possibility of being integrated into lab-on-a-chip devices. 8,9 In contrast, label-free methods typically utilize differences in physical properties such as cell size, 10 density, 11 cell adhesion, [12][13][14][15] and dielectric properties. [16][17][18] One potential drawback in label-free methods is that the physical difference is not high enough for efficient separation in many cases, which limits the widespread use of the label-free methods.…”

Preparative Separation of Proteins Using Centrifugal Precipitation Chromatography Based on Solubility in Ammonium Sulfate Solution

“…(Zeus Industrial Products, Raritan, NJ, USA) was fastened to an aluminum spinning plate by nylon ties through multiple holes made along a spiral path on the plate [ Fig. [7,8] A strip of dialysis tubing with MWCO of 3,500 (ca. This column geometry forms a radial gradient of the centrifugal force to stabilize the AS gradient in the sample channel.…”