Differentiation of canine bone marrow stromal cells into voltage- andglutamate-responsive neuron-like cells by basic fibroblast growth factor

Journal of Veterinary Medical Science

Ken OKABAYASHI³  Kenji TESHIMA¹  Hiroshi SUGIYA³  Kazuya EDAMURA¹  Takanori NARITA³  Tomohiro NAKAYAMA²  Rei NAKANO¹  Kazushi ASANO¹ 

[1] Laboratory of Veterinary Surgery, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252�?0880, Japan;Laboratory of Veterinary Radiology, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252�?0880, Japan;Laboratory of Veterinary Biochemistry, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252�?0880, Japan

关键词: basic fibroblast growth factor;    bone marrow stromal cell;    canine;    neuron;   

DOI  :  10.1292/jvms.14-0284

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(38)Cited-By(1)We investigated the invitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage-and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow ofhealthy beagle dogs. Canine BMSCs were incubated with the basal medium for neuronscontaining recombinant human basic fibroblast growth factor (bFGF; 100ng/ml). The viability of the bFGF-treated cells wasassessed by a trypan blue exclusion assay, and the morphology was monitored. Real-timeRT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glialmarkers. Western blotting and immunocytochemical analysis for the neuronal markers wereperformed to evaluate the protein expression and localization. The Ca2+mobilization of the cells was evaluated using the Ca2+ indicator Fluo3 tomonitor Ca2+ influx. To investigate the mechanism of bFGF-induced neuronaldifferentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in themaintenance of the viability of canine BMSCs for 10 days, in the expression of neuronalmarker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore,in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increasein intracellular Ca2+ levels. Each inhibitor significantly attenuated thebFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGFcontributes to the differentiation of canine BMSCs into voltage- and glutamate-responsiveneuron-like cells and may lead to the development of new cell-based treatments forneuronal diseases.

【 授权许可】

Unknown