Effect of Single Growth Factor and Growth Factor Combinations on Differentiation of Neural Stem Cells

OBJECTIVE: The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF), insulin growth factor-I (IGF-I), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. METHODS: Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using beta-tubulin III and NeuN as neural markers. RESULTS: Neural differentiations in the presence of individual growth factors for beta-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-1, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for beta-tubulin III and 3.1% for NeuN. For beta-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1%; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). CONCLUSION: Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and postmitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.

The Cell Survival and Differentiation after Transplantation, Which Harvest from Adult Rat Brain by High-speed Centrifugation Method

OBJECTIVE: Many recent reports have shown that the mature mammalian brain harbors multipotent stem cells, rendering the brain capable of generating new neurons and glia throughout life. Harvested stem cells from an adult rat are transplanted in order to evaluate the cell survival and differentiation. METHODS: Using a percoll gradient with a high speed centrifugation method, we isolate neural stem/progenitor cells were isolated from the subventricular zone(SVZ) of a syngeneic adult Fisher 344 rats brain. For 14days expansion, the cultured cells comprised of a heterogeneous population with the majority of cells expressing nestin and/or GFAP. After expanding the SVZ cells in the presence of basic fibroblast growth factor-2, and transplanting then into the hippocampus of normal rats, the survival and differentiation of those cells were examined. For transplantation, the cultured cells were labeled with BrdU two days prior to use. In order to test their survival, the cells were transplanted into the dorsal hippocampus of normal adult Fisher 344 rats. RESULTS: The preliminary data showed that at 7days after transplantation, BrdU+ transplanted cells were observed around the injection deposition sites. Immuno-fluorescent microscopy revealed that the cells co-expressed BrdU+ and neuronal marker beta-tubulin III. CONCLUSION: The data demonstrate that the in vitro expanded SVZ cells can survive in a heterotypic environment and develop a neuronal phenotype in the neurogenic region. However more research will be needed to examine the longer survival time points and quantifying the differentiation in the transplanted cells in an injured brain environment.

Management Options and Prognosis of Carotid Artery Occlusive Disease / 대한뇌혈관외과학회지

OBJECT: The effective management of carotid occlusive disease still remains a challenge to neurosurgeons. The authors analyzed the series of management of carotid occlusive disease in order to determine whether our management strategy affected patient's clinical outcomes. Methods of identifying patients who stand to benefit from this therapy need to be established. METHOD: Clinical findings, management, complications and outcome in 52 patients with high grade carotid stenosis of at least 70% and occlusion were investigated. The patients were treated by percutaneous transluminal angioplasty (PTA) and/or stent (PTAS), Extracranial-Intracranial (EC-IC) bypass surgery, carotid endarterectomy (CEA) according to the neurologic status, medical condition, severerity of stenosis, collateral blood flow. RESULTS: The causes of carotid stenosis were 40 atherosclerosis arteries, 9 spontaneous dissections and 2 traumas, 1 fibomuscular dysplasia (FMD) of 52 patients. 9 patients were treated by PTA alone, and 28 patients by PTAS, 9 patients by EC-IC bypass surgery, 3 patients by PTAS followed by EC-IC bypass surgery, 3 patients by CEA. For the outcome according to management, 26 patients (100% of all bypass surgery only and CEA, 35% of all PTA and PTAS) recovered excellently, 14 patients (35% of all PTA and PTAS) had a good outcome. 2 patients died. CONCLUSION: The results of this study suggest that PTAS should be a useful and effective treatment method for some patients with the severe atherosclerotic stenosis or carotid artery dissection. However, the surgical management must be considered for the high risk, high grade carotid stenosis patients with collateral blood flow, and with or without mild or moderate deficits.