A Prospective Study of Bone Marrow Hematopoietic and Mesenchymal Stem Cells in Type 1 Gaucher Disease Patients
Figure 2
(A) Relative number of CD34+ cells was measured in GD (n = 7) and controls (n = 10) BM samples according to ISHAGE Guidelines [34]. (B) CD34+ cells clonogenic potential. Granulocyte–macrophage colony-forming units (CFU-GM), erythroid burst-forming units (BFU-E), and megakaryocyte colony-forming units (CFU-Mk) were scored at day 14 by direct visualization using an inverted microscope (n = 8 and 10 for GD patients and controls, respectively). (C) BM-CD34+ cells ex-vivo expansion. Total nucleated cell (TNCs) number was determined after 6 and 12 days culture (n = 6 and 10 for GD and controls respectively). Results are presented as TNCs fold expansion. (D) Primitive hematopoiesis was evaluated by LTC-IC assay. GD patients and controls CD34+ cells were cultured on healthy donors MSCs during a period of 5 weeks. All experiments were performed in triplicate (n = 4 and 8 for GD patients and controls, respectively). (E) Four- to 8-week-old NSG mice were transplanted by retro-orbital injection with 1.105 BM CD34+ cells from GD patients or controls. The presence of human CD45+ cells was assessed by flow cytometry 7 weeks later (n = 5 mice engrafted with CD34+ cells from 2 healthy donors and 4 mice engrafted with CD34+ cells from 2 GD patients).