Bone marrow CD34⁺ cells and clonogenic progenitor cells in AIC patients.

The left bars represent the mean percentages (± SEM) of CD34⁺ cells in 15 AIC patients and 20 normal controls obtained in 2-color flow cytometric analysis of BMMCs. The right bars represent the mean colony values (± SEM) obtained from BMMCs in the clonogenic progenitor cell assays. We cultured 10⁵BMMCs from AIC patients (n = 13) and normal controls (n = 16) in 1 mL methylcellulose 0.9% in Iscove modified Dulbecco medium supplemented with 30% fetal calf serum (PAA Laboratories GmbH, Linz, Austria), 1% bovine serum albumin (BSA; Sigma, St Louis, MO), 10⁻⁴ mol/L mercaptoethanol (Sigma), 0.075% sodium bicarbonate (GibcoBRL; Life Technologies), and 2 mmol/L L-glutamine (Sigma), in the presence of 5 ng GM-CSF, 50 ng interleukin (IL)-3 and 2 IU erythropoietin for CFU-GM and BFU-E colony formation. Colonies were enumerated on day 14. We cultured 10⁶ BMMCs from AIC patients (n = 12) and normal controls (n = 10) in MegaCult-C medium for CFU-Meg colony growth. Colonies were scored after 10 to 12 days of incubation after fixation and staining by alkaline phosphatase antialkaline phosphatase technique with the use of anti-CD41 monoclonal antibody. Comparison between patients and normal controls was performed by means of the 2-tailed Student t test.