Proliferative capacity of the isolated single CD(34)(+) glycosylphosphatidylinesitol-anchored (GPI) protein negative and positive hematopoietic cells in paroxysmal nocturnal hemoglobinuria / 中华血液学杂志

<p><b>OBJECTIVES</b>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).</p><p><b>METHOD</b>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.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSION</b>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.</p>

Ex vivo expansion of CD34(+) CD59(+) cells from bone marrow of paroxysmal nocturnal hemoglobinuria patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the separation, purification and ex vivo expansion of CD(34)(+) CD(59)(+) cells from patients with paroxysmal nocturnal hemoglobinuria (PNH), and explore the new treatment for the PNH patients.</p><p><b>METHODS</b>CD(34)(+) CD(59)(+) cells were selected from the bone marrow mononuclear cells of PNH patients by means of immunomagnetic microbead-flow cytometry two step sorting method, followed by ex vivo expansion of the cells with combination of hematopoietic factors for two weeks.</p><p><b>RESULTS</b>The best combination for the ex vivo expansion was SCF + IL-3 + IL-6 + FL + Tpo + Epo, and the seventh day was the most suitable time for the best harvest when the CD(34)(+) CD(59)(+) cells were 22.42 +/- 3.73 fold expanded. After ex vivo expansion, the cells remained CD(59) positive and potent capacity of colony formation, but their potentialities to multilineage differentiation were decreased.</p><p><b>CONCLUSION</b>The present study shows that ex vivo expansion of CD(34)(+) CD(59)(+) cells from PNH patients might promise the possibility of performing ABMT or APBSCT clinincally for the patients.</p>