Bone marrow-derived mesenchymal stem cells differentiate into nerve-like cells in vitro after transfection with brain-derived neurotrophic factor gene

• Published in: In vitro cellular & developmental biology. Animal Vol. 51; no. 3; pp. 319 - 327
• Main Authors: Liu, Qianxu, Cheng, Guangui, Wang, Zhiwei, Zhan, Shujie, Xiong, Binbin, Zhao, Xiaoming
• Format: Journal Article
• Language: English
• Published: Boston Springer-Verlag 01.03.2015; Springer Science & Business Media LLC; Springer US; Society for In Vitro Biology
• Subjects: adults; Animal Genetics and Genomics; Animals; antigens; Biomedical and Life Sciences; bone marrow cells; Bone Marrow Cells - cytology; Brain-Derived Neurotrophic Factor - genetics; Brain-Derived Neurotrophic Factor - metabolism; Cell Biology; Cell Culture; Cell Cycle; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cell Proliferation; Cell Shape; Culture Media - chemistry; Developmental Biology; endothelial cells; enzyme-linked immunosorbent assay; erythrocytes; fibroblasts; Flow Cytometry; Gene Expression Regulation; genes; Genetic Vectors - metabolism; Guinea Pigs; HEK293 Cells; Humans; immunocytochemistry; Immunohistochemistry; Life Sciences; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; neurons; Neurons - cytology; Neurons - metabolism; Open Reading Frames - genetics; Phenotype; retinoic acid; reverse transcriptase polymerase chain reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; STEM CELLS; stromal cells; Transfection; Nerve-like cells; Flow cytometry; BMSC; Differentiation; gene
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-015-9875-1

Bone marrow-derived mesenchymal stem cells can differentiate into a variety of adult cells. Brain-derived neurotrophic factor (BDNF) is briefly active during differentiation and induces mesenchymal stem cells to differentiate into nerve cells. In this study, we cloned human BDNF to generate a recombinant pcDNA3.1(−)-BDNF vector and transfected the vector into bone marrow-derived mesenchymal stem cells. We selected these cells with Geneticin-418 to obtain BDNF-BMSCs, which were induced with retinoic acid to obtain induced BDNF-BMSCs. The transfected cells displayed the typical morphology and surface antigen profile of fibroblasts and were observed to express clusters of differentiation 29, 44, and 90 (observed in matrix and stromal cells), but not clusters of differentiation 31, 34, and 45 (observed in red blood cells and endothelial cells), via flow cytometry. Enzyme-linked immunosorbent assays showed that transfected bone marrow-derived mesenchymal stem cells secreted more BDNF than non-transfected bone marrow-derived mesenchymal stem cells. Immunocytochemistry and real-time reverse transcription polymerase chain reaction analysis showed that non-induced BDNF-BMSCs maintained a higher proliferative capacity and expressed higher amounts of brain-derived neurotrophic factor, nestin, neuron-specific enolase, and glial fibrillary acid protein than non-transfected bone marrow-derived mesenchymal stem cells. An additional increase was observed in the induced BDNF-BMSCs compared to the non-induced BDNF-BMSCs. This expression profile is characteristic of neurocytes. Our data demonstrate that bone marrow-derived mesenchymal stem cells transfected with the BDNF gene can differentiate into nerve-like cells in vitro, which may enable the generation of sufficient quantities of nerve-like cells for treatment of neuronal diseases.