Elutriation for isolation of megakaryocytes.
Successful isolation of guinea pig megakaryocytes in large numbers was first achieved with a combination of techniques, taking sequential advantage of the low relative densities and large diameters of most megakaryocytes. Several laboratories have made minor improvements, but this approach retains the disadvantage of losing a significant fraction of the megakaryocyte population, the small immature ones. Counterflow centrifugal elutriation has been shown to eject cells from a chamber progressively, according to their sizes. Because almost all the megakaryocytes are bigger than the other marrow cells, the megakaryocytes can be retained while rejecting the contaminants. With this technology, yields of 1.4-2.0 x 10(6) megakaryocytes from one guinea pig are routine, recoveries have been 93%-94% of the input number of megakaryocytes, and final purities now average 72%. A split-specimen comparison with our previous method found elutriation to provide much greater yield and recovery with at least as great a purification as the density-velocity combination. This new technique was easily adapted to isolation of megakaryocytes in single aspirates from normal human marrow. Fifty-fold purification with near total recovery and a yield of 27,000 megakaryocytes per donor allows easy and reliable cytologic studies. Elutriation appears to be the current method of choice for isolation of megakaryocytes.
Shoff
P K
PK
VA Medical Center, Washington, D.C. 20422.
Levine
R F
RF
D000818:Animals; D001854:Bone Marrow Cells; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D002498:Centrifugation / Q000295:instrumentation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D006168:Guinea Pigs; D008533:Megakaryocytes / Q000166:cytology*