STUDY ON INDUCTION AND DIFFERENTIATION OF NEURAL STEM CELLS DERIVED FROM ADULT HUMAN BONE MARROW / 解放军医学杂志

ABSTRACT

To explore the feasibility and validity of isolated neural stem cells(NSC) derived from adult human bone marrow, the adult human bone marrow stromal cells, harvested by density gradient centrifugation following an ilium puncture, were cultured in "Cytokines NSC medium" for ascertaining the optimal survival conditions in vitro . Proliferation of NSCs was evaluated by formation of cell clones. Antibodies against Nestin, NSE and GFAP were chosen for identifying NSCs, neurons and glial cells, respectively. The involved cytokines in culture included GDNF(20ng/ml), LIF(10ng/ml)and RA(0 5?g/ml). Among the adult human bone marrow stromal cells, there were some NSCs with rough and large cytoplasmic granules proliferating rapidly into islets shaped cellular spheres with positive Nestin, a specific antigen on the fetal neural epithelium. After separating the cellular spheres into single cells and then plating them, we found the islets shaped cellular spheres appeared again. Following differentiation of the NSCs spheres, some of them formed small buds,which then developed further into long projects connecting each other. Most of the cells with long projects showed positive NSE or GFAP. It is possible that adult human bone marrow stromal cells could be induced into NSCs under certain experimental conditions. Also it implied the feasibility and validity that the adult human bone marrow might be used as the seed cells of the neural stem cells.