Methods and compositions for separating cells
View Patent ↗The invention provides compositions and methods for cell separation. These reagents and techniques specifically agglutinate cells via surface antigen recognition and can be used to recover rare cell types in high yield.
1. A cell separation reagent comprising:
a) dextran;
b) anti-glycophorin A antibody;
c) heparin;
d) serum albumin; and
e) ethylenediaminetetraacetic acid (EDTA) wherein dextran, anti-glycophorin A antibody, heparin, serum albumin, and EDTA are in solution.
2. The reagent of claim 1 , further comprising phosphate buffered saline.
3. The reagent of claim 1 , wherein the pH of said reagent is between 6.8 to 7.8.
4. The reagent of claim 1 , wherein said serum albumin is bovine serum albumin.
5. The reagent of claim 1 , wherein said serum albumin is human serum albumin.
6. The reagent of claim 1 , wherein said anti-glycophorin A antibody is monoclonal.
7. The reagent of claim 1 , wherein said anti-glycophorin A antibody is an IgM antibody or an IgG antibody.
8. The reagent of claim 1 , wherein said anti-glycophorin A antibody is an anti-human glycophorin A antibody.
9. The reagent of claim 1 , wherein the concentration of said anti-glycophorin A antibody is about 0.001 mg/L to about 15 mg/L.
10. The reagent of claim 1 , wherein the concentration of said serum albumin is about 0.5% to about 5%.
11. The reagent of claim 1 , wherein the concentration of EDTA is 0.1 mM to 5 mM.
12. A kit comprising a blood collection vessel and the cell separation reagent of claim 1 .
13. The kit of claim 12 , wherein said blood collection vessel is a blood bag.
14. The kit of claim 12 , wherein said blood collection vessel is a vacuum tube.
15. A method for separating cells, said method comprising
a) contacting a blood cell-containing sample with the cell separation reagent of claim 1 ;
b) allowing said sample of step a) to partition into an agglutinate and a supernatant phase at 1 xg; and
c) recovering said cells from the agglutinate or the supernatant phase.
16. The method of claim 15 , wherein said sample is a human blood cell-containing sample.
17. The method of claim 15 , wherein said sample is a peripheral blood sample.
18. The method of claim 15 , wherein said sample is an umbilical cord sample.
19. The method of claim 15 , wherein said sample is a bone marrow sample.
20. The method of claim 15 , wherein said cells are recovered from said supernatant phase.