PMID- 25773996 OWN - NLM STAT- MEDLINE DCOM- 20151215 LR - 20220408 IS - 1543-706X (Electronic) IS - 1071-2690 (Print) IS - 1071-2690 (Linking) VI - 51 IP - 3 DP - 2015 Mar TI - Bone marrow-derived mesenchymal stem cells differentiate into nerve-like cells in vitro after transfection with brain-derived neurotrophic factor gene. PG - 319-27 LID - 10.1007/s11626-015-9875-1 [doi] AB - 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. FAU - Liu, Qianxu AU - Liu Q AD - Department of Otolaryngology, Zhuhai Hospital of Medical College of Jinan University, Zhuhai, 519000, People's Republic of China. FAU - Cheng, Guangui AU - Cheng G FAU - Wang, Zhiwei AU - Wang Z FAU - Zhan, Shujie AU - Zhan S FAU - Xiong, Binbin AU - Xiong B FAU - Zhao, Xiaoming AU - Zhao X LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150314 PL - Germany TA - In Vitro Cell Dev Biol Anim JT - In vitro cellular & developmental biology. Animal JID - 9418515 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Culture Media) RN - 0 (RNA, Messenger) RN - 7171WSG8A2 (BDNF protein, human) SB - IM MH - Animals MH - Bone Marrow Cells/*cytology MH - Brain-Derived Neurotrophic Factor/*genetics/metabolism MH - Cell Cycle MH - *Cell Differentiation/drug effects/genetics MH - Cell Proliferation MH - Cell Shape MH - Culture Media/chemistry MH - Flow Cytometry MH - Gene Expression Regulation MH - Genetic Vectors/metabolism MH - Guinea Pigs MH - HEK293 Cells MH - Humans MH - Immunohistochemistry MH - Mesenchymal Stem Cells/*cytology/metabolism MH - Neurons/*cytology/metabolism MH - Open Reading Frames/genetics MH - Phenotype MH - RNA, Messenger/genetics/metabolism MH - *Transfection PMC - PMC4368845 EDAT- 2015/03/17 06:00 MHDA- 2015/12/17 06:00 PMCR- 2015/03/14 CRDT- 2015/03/17 06:00 PHST- 2014/04/09 00:00 [received] PHST- 2015/01/26 00:00 [accepted] PHST- 2015/03/17 06:00 [entrez] PHST- 2015/03/17 06:00 [pubmed] PHST- 2015/12/17 06:00 [medline] PHST- 2015/03/14 00:00 [pmc-release] AID - 9875 [pii] AID - 10.1007/s11626-015-9875-1 [doi] PST - ppublish SO - In Vitro Cell Dev Biol Anim. 2015 Mar;51(3):319-27. doi: 10.1007/s11626-015-9875-1. Epub 2015 Mar 14.