[Proliferative capacity of the isolated single CD(34)(+) glycosylphosphatidylinesitol-anchored (GPI) protein negative and positive hematopoietic cells in paroxysmal nocturnal hemoglobinuria].
OBJECTIVE
To investigate the stroma-independent growth ability, multilineage differentiation and expansion of the single hematopoietic stem/progenitor cell from patients with paroxysmal nocturnal hematoglobinuria (PNH).


METHODS
The CD(34)(+) CD(59)(+) and CD(34)(+) CD(59)(-) cells from PNH patients and CD(34)(+) CD(59)(+) cells from normal volunteers were sorted as each single cell into a well of 96 well culture plates containing culture medium supplemented with SCF, IL-3, Epo, GM-CSF, G-CSF, IL-6, Tpo and Flt-3 ligand.


RESULTS
(1) Single PNH CD(34)(+) CD(59)(-) cell had a higher capacities for plating efficiency, colony (>/= 50 cells) formation and cell expansion than that of the PNH CD(34)(+) CD(59)(+) cells (P < 0.05); (2) Both the single CD(34)(+) CD(59)(-) cells from PNH patients and the single CD(34)(+) CD(59)(+) cells from normal controls had similar capacities for cell plating efficiency and colony and large colony formation. The PNH CD(34)(+) CD(59)(-) cells had a lower average cell production and cell expansion capacity. (3) The single CD(34)(+) CD(59)(+) cells from both PNH patients and normal controls showed the same capacities for cell plating efficiency and colony formation. The PNH CD(34)(+) CD(59)(+) cells exhibited much lower capacity for large colony formation, average cell production and total cell expansion. (4) A diminished secondary colony formation ability was also observed in the PNH CD(34)(+) CD(59)(+) and CD(34)(-) CD(59)(-) clones.


CONCLUSIONS
The single PNH CD(34)(+) CD(59)(-) cells had growth advantage over the single PNH CD(34)(+) CD(59)(+) cells to some extent, but they both had impaired growth abilities as compared with CD(34)(+) cells from normal volunteers.
Department of Hematology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China.
Bing
B
Han
Yongji
Y
Wu
Zhaojiang
Z
Lu
Zhinan
Z
Zhang
D018952:Antigens, CD34 / Q000276:immunology*; D018957:CD59 Antigens / Q000276:immunology; D018929:Cell Culture Techniques; D002455:Cell Division / Q000502:physiology; D003114:Colony-Forming Units Assay; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000473:pathology*; D006457:Hemoglobinuria, Paroxysmal / Q000503:physiopathology*; D006801:Humans
D018952:Antigens, CD34; D018957:CD59 Antigens