Study On Conversion Of Adult Human Myoblasts Into Neural Precursor Cells And Transplantation For Focal Cerebral Ischemia In Rats

Objective To investigate the feasibility of inducing adult human myoblasts into neural precursor cells in vitro and whether neural precursor cells derived from myoblasts could be used for cell transplantation in experimental cerebral ischemia and be contributed to the treatment of stroke. And to investigate whether myoblast-derived neural precursor cells delivered to the carotid artery could enter the ischemic brain and find new way of these cells transplantation into the brain.Methods The study consisted of three parts. Part â…  The myoblasts were harvested and isolated from minced adult human temporal muscle samples. After purified clonally, they were incubated with no-serum medium including basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and leukemia inhibitory factor (LIF) in vitro. Morphological change was investigated during incubation period. Bromodeoxyuridine (BrdU) was used to evaluate cell proliferation. Immunofluorescence cytochemistry and RT-PCR analysis for nestin, neuronal (neurofilament68, NF68), astrocytic (Glial fibrillary acidic protein, GFAP), oligodendrocytic (Galactocerebroside, Galc) markers were used to identify the differentiation. Part â…¡ A stroke model in rats with photochemically induced thrombosis of proximal cerebral middle artery (MCA) was introduced for cell transplantation. Neural precursor cells derived from myoblasts were transplanted into the area near the ischemic cortex. Each rat was subjected to a series of behavioral tests (tactile stimulation test, limb placement test and equilibrium test) to evaluate neurologic function before and 7, 14, 21 and 28 days after stroke. Immunohistochemistry was used to identify cell-specific proteins (GFAP, MAP2, Galc) of transplanted cells identified by human nuclear specific antibody. The survival, migration and differentiation of human cells were evaluated. Hematoxylin-eosin staining and imaging analysis system were used to assess lesion volume. Part â…¢ Rats were subjected transient (90 min) middle cerebral artery occlusion and treated with intra carotid arterial injection of myoblasts and myoblast-derived neural precursor cells 24 h after ischemia. As in Part â…¡, immunohistochemistry was used to evaluate the survival, migration and differentiation of transplanted cells.Results Part â…  After induction, cells became non-adherent aggregates as...