Purification of human megakaryocytes using a glycoprotein Ib dependent agglutination technique.
In order to study normal and abnormal human megakaryocytopoiesis we modified a glycoprotein Ib dependent megakaryocyte purification technique originally described in guinea pigs. After Percoll density centrifugation, 2 ml bone marrow aspirates were subjected to 3 steps of bovine plasma induced cell clumping, which should selectively agglutinate glycoprotein Ib positive cells. In 13 experiments the resulting samples contained 28-102 x 10(3) megakaryocytes (median 70) with a median purity of 79%. Megakaryocyte recovery ranged from 30-86% (median 51%). The DNA profile showed median values of 10% for 8 N, 44% for 16 N and 43% for 32 N megakaryocytes. The described method is relatively simple, inexpensive and gives results comparable with more elaborate techniques.
van Pampus
E C
EC
Department of Haematology, Free University Hospital, Amsterdam, The Netherlands.
Huijgens
P C
PC
Wijermans
P W
PW
Ossenkoppele
G J
GJ
Langenhuijsen
M M
MM
D000371:Agglutination / Q000276:immunology; D002452:Cell Count / Q000379:methods; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D004247:DNA / Q000032:analysis; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology*; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*
D010980:Platelet Membrane Glycoproteins; D004247:DNA