[Study on the expansion of megakaryocyte progenitors in vitro from cord blood].
This study was aimed to investigate the effect of various cytokines on megakeryocytes expansion in vitro from human cord blood CD34(+) cells in order to establish an optimal culture system for MK expansion. Mononuclear cells were obtained by Ficoll-Hapaque density gradient separation. CD34(+) cells were positively isolated using a CD34 progenitor cell isolation kit. CD34(+) cells were placed into 24 well plates at a concentration of 2 x 10(4) per well. Each well contained 1 ml of IMDM with the present of effective MK cells growth cytokines. Clonogenic potentials of MK progenitor were assayed using a methylcellulose cultures system. The results suggested that four cytokines (IL-3 + IL-6 + TPO + FLT3L) culture system could effectively induce and expand cord blood CD41(+) MK cells. The number of CD41(+) cells expanded 154.67 +/- 32.21-fold on day 7, and 193.23 +/- 25.24-fold on day 14. In conclusion, established expansion system in vitro for MK cells provides experimental foundation for recovery of platelets after cord blood transplantation.
Chen
Guo-Hua
GH
Department of Pediatrics, Huizhou Central Hospital, Huizhou, China.
Fang
Jian-Pei
JP
Xu
Hong-Gui
HG
Liu
Si-Xi
SX
Huang
Shao-Liang
SL
D018952:Antigens, CD34 / Q000032:analysis; D001792:Blood Platelets / Q000166:cytology / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D008533:Megakaryocytes / Q000166:cytology* / Q000276:immunology; D008565:Membrane Proteins / Q000494:pharmacology; D040201:Platelet Membrane Glycoprotein IIb / Q000032:analysis; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology; D013926:Thrombopoietin / Q000494:pharmacology
D018952:Antigens, CD34; D007377:Interleukin-3; D015850:Interleukin-6; D008565:Membrane Proteins; D040201:Platelet Membrane Glycoprotein IIb; C084881:flt3 ligand protein; D013926:Thrombopoietin