Specific induction of neuronal cells from bone marrow stromal cells and application for autologous transplantation
Mari Dezawa, … , Yoshihisa Suzuki, Chizuka Ide
Mari Dezawa, … , Yoshihisa Suzuki, Chizuka Ide
Published June 15, 2004
Citation Information: J Clin Invest. 2004;113(12):1701-1710. https://doi.org/10.1172/JCI20935.
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Article Neuroscience

Specific induction of neuronal cells from bone marrow stromal cells and application for autologous transplantation

Abstract

Bone marrow stromal cells (MSCs) have the capability under specific conditions of differentiating into various cell types such as osteocytes, chondrocytes, and adipocytes. Here we demonstrate a highly efficient and specific induction of cells with neuronal characteristics, without glial differentiation, from both rat and human MSCs using gene transfection with Notch intracellular domain (NICD) and subsequent treatment with bFGF, forskolin, and ciliary neurotrophic factor. MSCs expressed markers related to neural stem cells after transfection with NICD, and subsequent trophic factor administration induced neuronal cells. Some of them showed voltage-gated fast sodium and delayed rectifier potassium currents and action potentials compatible with characteristics of functional neurons. Further treatment of the induced neuronal cells with glial cell line–derived neurotrophic factor (GDNF) increased the proportion of tyrosine hydroxylase–positive and dopamine-producing cells. Transplantation of these GDNF-treated cells showed improvement in apomorphine-induced rotational behavior and adjusting step and paw-reaching tests following intrastriatal implantation in a 6-hydroxy dopamine rat model of Parkinson disease. This study shows that a population of neuronal cells can be specifically generated from MSCs and that induced cells may allow for a neuroreconstructive approach.

Authors

Mari Dezawa, Hiroshi Kanno, Mikio Hoshino, Hirotomi Cho, Naoya Matsumoto, Yutaka Itokazu, Nobuyoshi Tajima, Hitoshi Yamada, Hajime Sawada, Hiroto Ishikawa, Toshirou Mimura, Masaaki Kitada, Yoshihisa Suzuki, Chizuka Ide

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Analysis of TF-MSCs (5 days after trophic factor induction). (A–C) Phase...
Analysis of TF-MSCs (5 days after trophic factor induction). (A–C) Phase contrast of TF-MSCs from rats (A and B) and humans (C). Bars, A = 200 ∝m, B and C = 50 ∝m. (D–F and H–J) Immunocytochemical analysis of neuronal and glial markers in rat (F, H–J) and human (D and E) TF-MSCs. MAP-2ab (D), neurofilament-M (E), and β-tubulin isotype 3 (F) were detected. None of the cells were reactive to GFAP (H), GalC (I), and O4 (J). (G) The Brd-U labeling of rat TF-MSCs. MAP-2ab–positive cells (green) did not incorporate Brd-U (red), whereas negative cells were occasionally incorporated with Brd-U. Bars in D–J = 100 ∝m. (K) Western blot analysis of MAP-2ab and GFAP rat samples. Brain, positive control; TF-MSC. β-tubulin (tub) as a loading control. (L–Q) Patch clamp. K+ current increased with trophic factor induction up to approximately 1,600 pA and 4,000 pA in rat (L) and human (M) TF-MSCs, respectively. (N) Phase contrast of human TF-MSCs recorded in (M). (O) Voltage-gated inward current recorded in rat BDNF + NGF–treated TF-MSCs. A series of Na current to show the process of block by TTX. Capacity current was blanked. (P) Action potentials from rat BDNF + NGF–treated TF-MSCs; subthreshold, threshold, and suprathreshold current injections were made. (Q) Immunocytochemistry of sodium channel (green). Bar = 30 –m. (R) Relative promoter activities of NeuroD and GFAP in rat MSCs, N-MSCs, and TF-MSCs.