Biological characteristics of neural stem cells derived from adult bone marrow by immunomagnetic beads / 中国组织工程研究

ABSTRACT

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) under certain conditions can differentiate into a variety of connective tissue cells, also can be turned into neurons and glial cells. OBJECTIVE: To investigate biological characteristics of neural stem cells (NSCs) from adult bone marrow by immunomagnetic beads. DESIGN, TIME AND SETTING: Autologous bone marrow was selected as observational objects. Experiments were performed at the Department of Clinical Laboratory, Heze Municipal Hospital from March 2003 to June 2007. MATERIALS: Bone marrow was collected from inpatients with stroke, senile dementia, Parkinson's disease and cerebral ischemia at the Department of Neurology, Heze Municipal Hospital. METHODS: NSCs were harvested by immunomagnetic beads through isolating immunomagnetic beads from other cells under magnetic field following combination between specific antibody of immunomagnetic bead surface and NSC antigen in bone marrow. Purified NSCs were inoculated in Eagles MEM medium containing fetal bovine serum (FBS) of 0.20 volume fraction. Primary culture was performed in the medium in addition of basic fibroblast growth factors (bFGF) and epidermal growth factor (EGF). MAIN OUTCOME MEASURES: Immunohistochemistry and immunofluorescence were applied to identify the cells. Flow cytometry was used to measure DNA content. RESULTS: Under a phase contrast microscope, NSCs showed turgor vitalis, clear nucleus and nucleolus, enlarged nervous process and dense lattice. Immunohistochemistry and immunofluorescence demonstrated that neuron-specific enolase (NSE), neuroprotein, glial fibrillary acidic protein (GFAP) and microtubule-associated protein (MAP) were positive. Flow cytometry confirmed that DNA content was normal diploid, no induced mutation of the passaged cells. CONCLUSION: NSCs are harvested from adult bone marrow by combination between specific antibody of immunomagnetic bead surface and NSC antigen in bone marrow. NSCs have the characteristics of nerve cells in neurochemistry and morphology.