A novel serum free system for cloning human megakaryocytic progenitors (CFU-Meg). The role of thrombopoietin and other cytokines on bone marrow and cord blood CFU-Meg growth under serum free conditions.

摘要: Human megakaryocytic progenitors (CFU-Megs) are usually cloned in plasma clot cultures. Since the medium employed to prepare contains animal or human serum, there exists a potential risk that CFU-Megs growing vitro could be exposed serum derived megakaryopoietic inhibitors. To address this issue, we aimed establish relatively simple "serum free" cloning model for these progenitors. Accordingly, found if bone marrow cord blood CD34+ cells plated methylcellulose containing substitute, and stimulated with recombinant thrombopoietin (TpO), they exclusively form CFU-Meg colonies. Subsequently colonies can easily scored an inverted microscope based only on their morphological criteria. We efficiency of was higher our free system than traditional proposed paper is simple, moreover does not require time consuming immunostaining identify colonies, it should widely recommended studying megakaryopoiesis. also found, under conditions TpO crucial formation. In absence TpO, neither gp 130 activating cytokines (IL-6, IL-11, LIF, CNTF) nor other hematopoietic growth factors (KL, IL-3, GM-CSF, EpO) were able, when added alone, stimulate Finally, report enriched progenitors, moreover, from possess proliferative capacity isolated normal adult marrow.