Microfluidic Device For Cell Separation And Uses Thereof

Abstract

Methods for separating cells from a sample (e.g., separating fetal red blood cells from maternal blood) include introducing a sample including cells into one or more microfluidic channels. In one embodiment, the device includes at least two processing steps. For example, a mixture of cells is introduced into a microfluidic channel that selectively allows the passage of a desired type of cell, and the population of cells enriched in the desired type is then introduced into a second microfluidic channel that allows the passage of the desired cell to produce a population of cells further enriched in the desired type. The selection of cells is based on a property of the cells in the mixture, for example, size, shape, deformability, surface characteristics (e.g., cell surface receptors or antigens and membrane permeability), or intracellular properties (e.g., expression of a particular enzyme).

What is claimed is: 
     
         1 - 34 . (canceled) 
     
     
         35 . A method of collecting fetal red blood cells from a sample comprising maternal blood, the method comprising
 introducing a sample of maternal blood into a lysis device containing a lysis reagent to selectively lyse maternal blood cells, to provide a sample enriched in fetal blood cells compared to maternal blood cells; and   introducing the sample enriched in fetal blood cells into a cell binding device comprising an array of obstacles coated with:
 (i) binding moieties that bind selectively to maternal blood cells to produce a sample further enriched in fetal blood cells, or 
 (ii) binding moieties that bind selectively to fetal blood cells to produce a sample depleted of fetal blood cells; and 
   collecting fetal cells from:
 (i) the sample further enriched in fetal blood cells, or 
 (ii) the obstacles in the cell binding device to which the fetal blood cells have bound. 
   
     
     
         36 . The method of  claim 35 , wherein the lysis reagent selective lyses maternal red blood cells and comprises NH 4 Cl (0 to 150 mM)+NaHCO 3  (0.001 to 0.3 mM)+acetazolamide (0.1 to 100 μM)). 
     
     
         37 . The method of  claim 35 , wherein the cell binding device comprises an array of obstacles coated with binding moieties that bind selectively to maternal blood cells to produce a sample further enriched in fetal blood cells. 
     
     
         38 . The method of  claim 37 , wherein the binding moieties are antibodies. 
     
     
         39 . The method of  claim 37 , wherein the binding moieties comprise anti-CD45 antibodies that selectively bind to adult white blood cells. 
     
     
         40 . The method of  claim 35 , wherein the cell binding device comprises an array of obstacles coated with binding moieties that bind selectively to fetal blood cells to produce a sample depleted of fetal blood cells. 
     
     
         41 . The method of  claim 40 , wherein the binding moieties are antibodies. 
     
     
         42 . The method of  claim 40 , wherein the binding moieties bind to fetal hemoglobin transferring receptor (CD71), thrombospondin receptor (CD36), or glycophorin A (GPA), which selectively bind to fetal blood cells. 
     
     
         43 . The method of  claim 42 , wherein the binding moieties comprise anti-CD71 antibodies that selectively bind to fetal nucleated red blood cells. 
     
     
         44 . The method of  claim 35 , wherein the fetal cells are collected from the sample further enriched in fetal blood cells. 
     
     
         45 . The method of  claim 44 , further comprising analyzing the fetal cells. 
     
     
         46 . The method of  claim 35 , wherein the fetal cells are collected from the obstacles in the cell binding device to which the fetal blood cells have bound. 
     
     
         47 . The method of  claim 46 , further comprising analyzing the fetal cells.