Induction of Human Mesenchymal Stem Cells into Dopamine-Producing Cells with Different Differentiation Protocols

Abstract

Several reports have shown that human mesenchymal stem cells (MSCs) are capable of differentiating outside the mesenchymal lineage. We sought to induce MSCs to differentiate into dopamine-producing cells for potential use in autologous transplantation in patients with Parkinson's disease (PD). Following cell culture with various combinations of differentiation agents under serum-free defined conditions, different levels of up-regulation were observed in the protein expression of tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis. Further analysis of selected differentiation protocols revealed that the induced cells displayed a neuron-like morphology and expressed markers suggesting neuronal differentiation. In addition, there was an increase in Nurr 1, the dopaminergic transcription factor gene, concomitant with a decrease gamma-aminobutyric acid (GABA)ergic marker expression, suggesting a specific dopaminergic direction. Moreover, the induced cells secreted dopamine in response to depolarization. These results demonstrate the great therapeutic potential of human MSCs in PD.

Advances in the Combined Use of Adult Cell Therapy and Scaffolds for Brain Tissue Engineering

“…In this regard, another study demonstrated the importance of factors such as stem cell factor (SCF), produced in situ in the lesioned striatum, to promote the migration and engraftment of MSCs via SCF receptor c-kit (Bantubungi et al, 2008). In the context of PD, several teams, including ours, reported the neuronal differentiation of human MSCs toward a dopaminergic phenotype in vitro, indicating that these cells may constitute an alternative dopamine secreting source of cells (Trzaska & Rameshwar, 2011;Barzilay et al, 2008;Trzaska et al, 2007;). An interesting study also reported no major differences between MSCs from normal patients compared to MSCs isolated from parkinsonian patients, which may be induced to produce up to 30 % of dopaminergic neurons in vitro .…”

Mesenchymal Stem Cell-Based Therapies for Parkinson’s Disease: Progress, Controversies and Lessons for the Future

“…At present, there are a limited number of reports describing dopaminergic neuronal differentiation of MSCs from rodents [24,62,81,216,217] and humans [25,74,81,101,103,214,[218][219][220]. The majority of these studies employed the addition of extrinsic factors to cell cultures, such as cytokines and small molecules; however, the effects of gene transfection, conditioned medium and co-culturing, as well as combinations of these methods were also investigated.…”

“…Application of extrinsic factors alone has also resulted in the induction of MSCs toward a dopaminergic neuronal phenotype [25,62,74,103,216,217,219,220]. An NSC-like population was obtained from hMSCs through the formation of neurosphere-like structures in low-attachment culture flasks in the presence of serum-free medium containing EGF and FGF-2 [25].…”

“…Human MSCs 30% TH-positive (flow cytometry); DA secretion in response to depolarization Barzilay et al [219] Multi-stage:…”

“…The remaining studies employed similar procedures including: multiple-stage application of cytokines FGF-2 (1 week), SHH+FGF-8 (1 week) followed by BDNF (1 week) [217]; treatment with FGF-2/ EGF for 48 hours as step 1, then application of BHA, dbcAMP, IBMX, RA and GDNF for 96 hours as step 2 [103], and a later study by the same group found improved TH expression with addition of dbcAMP, IBMX and AA with BDNF alone or GDNF/RA/TGF-β3 in the second step [219]; application of CoCl 2 and Y-27632, to presumably cause hypoxia-inducible factor 1 activation and Rho kinase inhibition [216]; and treatment with cytokines FGF-2 and GDNF, and 10% fetal calf serum, for 2 weeks [220]. A varying degree of success was achieved, including depolarization-induced secretion of DA [103,219,220], and reduction of behavioral deficits in a Parkinsonian rat model [217].…”