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Year : 2018  |  Volume : 4  |  Issue : 3  |  Page : 139-141

Neurotrophic factor-based strategies to enhance survival and differentiation of neural progenitor cells toward the dopaminergic phenotype

Department of Neurosurgery, Neurocenter and Regenerative Neuroscience Cluster, Inselspital, University of Bern, CH-3010 Bern, Switzerland

Date of Submission24-Jul-2018
Date of Acceptance12-Sep-2018
Date of Web Publication09-Oct-2018

Correspondence Address:
Dr. Hans R Widmer
Department of Neurosurgery, Neurocenter and Regenerative Neuroscience Cluster, Inselspital, University of Bern, CH-3010, Bern
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/bc.bc_23_18

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Parkinson's disease (PD) is a neurodegenerative disorder that presents with hallmark clinical symptoms of tremor at rest, bradykinesia, and muscle rigidity. Stem cell therapy has emerged as an experimental treatment for PD. However, optimizing the cell culture condition that allows enhanced survival and differentiation of cells toward the dopaminergic phenotype remains a logistical challenge. Here, we discuss the utility of a combination of neurotrophin-4/5 (NT-4/5) and glial cell line-derived neurotrophic factor (GDNF) in increasing the dopaminergic phenotypic expression of rat ventral mesencephalic (VM) tissue. Using organotypic explant cultures of fetal human ventral mesencephalon, we observed that NT-4/5 and GDNF as single factors, or in combination on DAergic neurons, increased survival and number of tyrosine hydroxylase immunoreactive neurons as well as the dopamine content in the culture medium. The application of specific neurotrophic factors, such as NT-4/5 and GDNF, as cell culture supplements or as adjunctive therapy to cell transplantation may achieve improved functional outcomes when contemplating cell-based regenerative medicine for PD.

Keywords: Dopaminergic neurons, glial cell line-derived neurotrophic factor, neurotrophin-4/5, organotypic explant cultures, Parkinson's disease

How to cite this article:
Di Santo S, Widmer HR. Neurotrophic factor-based strategies to enhance survival and differentiation of neural progenitor cells toward the dopaminergic phenotype. Brain Circ 2018;4:139-41

How to cite this URL:
Di Santo S, Widmer HR. Neurotrophic factor-based strategies to enhance survival and differentiation of neural progenitor cells toward the dopaminergic phenotype. Brain Circ [serial online] 2018 [cited 2023 Jun 6];4:139-41. Available from: http://www.braincirculation.org/text.asp?2018/4/3/139/242912

  Introduction Top

Parkinson's disease (PD) is characterized by loss of dopaminergic (DAergic) neurons in the substantia nigra pars compacta. Drugs therapies are currently available for the treatment of PD, however long-term pharmacological treatment is often accompanied by serious side effects. Stem cell therapy has been suggested as potent treatment for PD because they may represent as robust biological source of dopamine.

  Stem Cell Therapy for Parkinson's Disease Top

It has been demonstrated that transplantation of human fetal nigral tissue is safe and may reinnervate the dopamine-depleted striatum in PD patients.[1],[2],[3],[4] However, the survival of DAergic neurons limits the efficacy of this transplant strategy.[1] In this regard, nonfetal tissue sources of dopamine have been examined in an attempt to increase DAergic survival; along this line of investigation, induced pluripotent stem cells and embryonic stem cells have been evaluated as rich sources of DAergic neurons, but their potential to restore the striatum function is still under investigation.[5] Another interesting approach to increase the survival of DAergic neurons either in culture or following transplantation could be the use of neurotrophic factors. In this context, glial cell line-derived neurotrophic factor (GDNF) and neurotrophin-4/5 (NT-4/5) support the improved growth and survival of DAergic neurons.[6] Neurotrophic signaling pathways may be involved in these observed cell-surviving effects. In particular, GDNF is a member of the transforming growth factor-beta superfamily and promotes DAergic survival and differentiation by activating a multicomponent receptor complex called RET and the GDNF family receptor. Interestingly, GDNF increased high-affinity dopamine uptake in cultures of the fetal midbrain, improving DAergic viability and stimulating differentiation.[7],[8] On the other hand, NT-4/5 belongs to the NT family and triggers a signaling pathway that involves the rat sarcoma-phosphatidylinositol 3-kinase-Protein Kinase B (Ras-PI3K-Akt) and the phospholipase C-gamma 1.[9],[10] NT-4/5 can regulate the morphology and improve the survival of DAergic neurons in mesencephalic primary cultures.[11],[12] The administration of GDNF and NT-4/5 increased the survival of rat ventral mesencephalic (VM) tyrosine hydroxylase immunoreactive (TH-ir) neurons along with stimulation of dopamine (DA) release in free-floating roller-tube (FFRT) cultures.[6] In addition, the ability of donor tissue storage in FFRT cultures supports the strategy to pretreat the cells with growth factors. Of note, in this respect, DAergic viability and functions have been restored in a rat model of PD by using fibroblast growth factor 2-mediated pregrafting expansion of primary VM precursor cells.[13],[14] To date, most studies have only explored the effects of monotherapy of neurotrophic factors on DAergic cell survival. Here, we discuss our experiments evaluating the therapeutic potential of combined GDNF and NT-4/5 administration on VM tissue of human origin in an attempt to reveal the application of neurotrophic factors as cell culture supplement or as an adjunct therapy for cell transplantation in PD.

  Neurotrophic Factor Treatment of Neural Progenitor Cells Top

We assessed the survival and differentiation potential of organotypic explants of the fetal human VM when cultured with or without GDNF and NT-4/5 singly or combined pretreatment. The combined pretreatment increased both cell number and DA content of TH-ir neurons better than using singular treatments of either neurotrophic factors alone. In addition, no difference is observed in culture volumes, while the level of lactate dehydrogenase in culture medium was decreased in all the treatment conditions. These findings advance our current knowledge on the contribution of neurotrophic factors for DAergic neurons in animal models.[6] Indeed, it has been demonstrated that the GDNF reduces apoptosis and stimulates DAergic fiber growth in DAergic neurons and fetal nigral grafts, respectively.[15],[16] Moreover, GDNF and NT-4/5 reduce oxidative stress-induced cell death, which is implicated in PD and other neurodegenerative diseases, through an anti-apoptotic mechanism.[17],[18] The reduction of apoptosis might be one of the events underlying the observed NT-4/5 and GDNF protective action in TH-ir neurons. In addition, the decrease of lactic acid dehydrogenase (LDH) levels after GDNF and NT-4/5 treatment offer an alternative mechanism. That LDH levels are not lower in the GDNF and NT-4/5 combined treatment than the single neurotrophic factor treatment suggests that the synergistic action of GDNF and NT-4/5 is not a direct action on cell death reduction, but rather an increase of maturation and/or differentiation of the TH-ir cells. Indeed, no significant variation in culture volume is observed between the different experimental conditions. In addition, the treatment with neurotrophic factors has no influence on the protein GFAP expression levels, as shown previously.[6] In contrast, this neurotrophic factor treatment could promote the survival and the growth of other neuronal cells including striatal and cortical GABAergic neurons,[9],[19],[20],[21] suggesting the preferential effects of GDNF and NT-4/5 on the neuronal phenotype.

Several lines of investigation suggest that the pretreatment of DAergic neurons with neurotrophic factors may be a potential strategy for the PD treatment. In this context, we showed that both number of cultured DAergic neurons and DA levels increased after BDNF treatment.[22] In addition, the combined action of GDNF and BDNF promoted the survival of rat fetal nigral tissue.[16] There remain some discrepant reports on the effects of neurotrophic factors on cell survival and differentiation. For example, dopamine levels may correlate with the TH-ir cell number even though no growth factor treatment was applied; in contrast, a prominent increase in dopamine levels is achieved with the combined GDNF and BDNF treatment compared to the use of these neurotrophic factors individually.[16],[23] Notably, a clinical study reported increased uptake of fluorodopa in 2 PD patients when graft is exposed to GDNF.[24] Further investigations are needed to better understand the benefit of GDNF and NT4/5 pretreatment in cultured cells of potential DAergic graft donors, as well as the effects of these neurotrophic factors as adjunct treatments in clinically relevant animal models of PD.

  Conclusion Top

In conclusion, these findings on combined pretreatment with neurotrophic factors of DAergic neurons support the potential of this strategy for enhancing the survival and differentiation of neural progenitor cells as graft source for transplantation therapy in PD. Adjunctive use of neurotrophic factors with cell therapy may also reveal improved functional outcomes.

Financial support and sponsorship

This research was financially supported by the HANELA Foundation, Switzerland, and the Swiss National Science Foundation.

Conflicts of interest

There are no conflicts of interest.

  References Top

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Hauser RA, Freeman TB, Snow BJ, Nauert M, Gauger L, Kordower JH, et al. Long-term evaluation of bilateral fetal nigral transplantation in Parkinson disease. Arch Neurol 1999;56:179-87.  Back to cited text no. 2
Hallett PJ, Cooper O, Sadi D, Robertson H, Mendez I, Isacson O, et al. Long-term health of dopaminergic neuron transplants in Parkinson's disease patients. Cell Rep 2014;7:1755-61.  Back to cited text no. 3
Kordower JH, Goetz CG, Chu Y, Halliday GM, Nicholson DA, Musial TF, et al. Robust graft survival and normalized dopaminergic innervation do not obligate recovery in a Parkinson disease patient. Ann Neurol 2017;81:46-57.  Back to cited text no. 4
Kriks S, Shim JW, Piao J, Ganat YM, Wakeman DR, Xie Z, et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson's disease. Nature 2011;480:547-51.  Back to cited text no. 5
Meyer M, Matarredona ER, Seiler RW, Zimmer J, Widmer HR. Additive effect of glial cell line-derived neurotrophic factor and neurotrophin-4/5 on rat fetal nigral explant cultures. Neuroscience 2001;108:273-84.  Back to cited text no. 6
Lin LF, Doherty DH, Lile JD, Bektesh S, Collins F. GDNF: A glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science 1993;260:1130-2.  Back to cited text no. 7
Widmer HR, Schaller B, Meyer M, Seiler RW. Glial cell line-derived neurotrophic factor stimulates the morphological differentiation of cultured ventral mesencephalic calbindin- and calretinin-expressing neurons. Exp Neurol 2000;164:71-81.  Back to cited text no. 8
Widmer HR, Hefti F. Neurotrophin-4/5 promotes survival and differentiation of rat striatal neurons developing in culture. Eur J Neurosci 1994;6:1669-79.  Back to cited text no. 9
Gupta VK, You Y, Gupta VB, Klistorner A, Graham SL. TrkB receptor signalling: Implications in neurodegenerative, psychiatric and proliferative disorders. Int J Mol Sci 2013;14:10122-42.  Back to cited text no. 10
Hynes MA, Poulsen K, Armanini M, Berkemeier L, Phillips H, Rosenthal A. Neurotrophin-4/5 is a survival factor for embryonic midbrain dopaminergic neurons in enriched cultures. J Neurosci Res 1994;37:144-54.  Back to cited text no. 11
Studer L, Spenger C, Seiler RW, Altar CA, Lindsay RM, Hyman C. Comparison of the effects of the neurotrophins on the morphological structure of dopaminergic neurons in cultures of rat substantia nigra. Eur J Neurosci 1995;7:223-33.  Back to cited text no. 12
Andereggen L, Meyer M, Guzman R, Ducray AD, Widmer HR. Effects of GDNF pretreatment on function and survival of transplanted fetal ventral mesencephalic cells in the 6-OHDA rat model of Parkinson's disease. Brain Res 2009;1276:39-49.  Back to cited text no. 13
Jensen P, Pedersen EG, Zimmer J, Widmer HR, Meyer M. Functional effect of FGF2- and FGF8-expanded ventral mesencephalic precursor cells in a rat model of Parkinson's disease. Brain Res 2008;1218:13-20.  Back to cited text no. 14
Zawada WM, Zastrow DJ, Clarkson ED, Adams FS, Bell KP, Freed CR. Growth factors improve immediate survival of embryonic dopamine neurons after transplantation into rats. Brain Res 1998;786:96-103.  Back to cited text no. 15
Sautter J, Meyer M, Spenger C, Seiler RW, Widmer HR. Effects of combined BDNF and GDNF treatment on cultured dopaminergic midbrain neurons. Neuroreport 1998;9:1093-6.  Back to cited text no. 16
Lingor P, Unsicker K, Krieglstein K. GDNF and NT-4 protect midbrain dopaminergic neurons from toxic damage by iron and nitric oxide. Exp Neurol 2000;163:55-62.  Back to cited text no. 17
Burke RE, Antonelli M, Sulzer D. Glial cell line-derived neurotrophic growth factor inhibits apoptotic death of postnatal substantia nigra dopamine neurons in primary culture. J Neurochem 1998;71:517-25.  Back to cited text no. 18
Widmer HR, Hefti F. Stimulation of GABAergic neuron differentiation by NT-4/5 in cultures of rat cerebral cortex. Brain Res Dev Brain Res 1994;80:279-84.  Back to cited text no. 19
Ventimiglia R, Mather PE, Jones BE, Lindsay RM. The neurotrophins BDNF, NT-3 and NT-4/5 promote survival and morphological and biochemical differentiation of striatal neurons in vitro. Eur J Neurosci 1995;7:213-22.  Back to cited text no. 20
Allen SJ, Watson JJ, Shoemark DK, Barua NU, Patel NK. GDNF, NGF and BDNF as therapeutic options for neurodegeneration. Pharmacol Ther 2013;138:155-75.  Back to cited text no. 21
Höglinger GU, Sautter J, Meyer M, Spenger C, Seiler RW, Oertel WH, et al. Rat fetal ventral mesencephalon grown as solid tissue cultures: Influence of culture time and BDNF treatment on dopamine neuron survival and function. Brain Res 1998;813:313-22.  Back to cited text no. 22
Studer L, Psylla M, Bühler B, Evtouchenko L, Vouga CM, Leenders KL, et al. Noninvasive dopamine determination by reversed phase HPLC in the medium of free-floating roller tube cultures of rat fetal ventral mesencephalon: A tool to assess dopaminergic tissue prior to grafting. Brain Res Bull 1996;41:143-50.  Back to cited text no. 23
Mendez I, Dagher A, Hong M, Hebb A, Gaudet P, Law A, et al. Enhancement of survival of stored dopaminergic cells and promotion of graft survival by exposure of human fetal nigral tissue to glial cell line – Derived neurotrophic factor in patients with Parkinson's disease. Report of two cases and technical considerations. J Neurosurg 2000;92:863-9.  Back to cited text no. 24

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