Journal of Molecular Neuroscience

, Volume 51, Issue 2, pp 307–317 | Cite as

A New Experimental Model for Neuronal and Glial Differentiation Using Stem Cells Derived from Human Exfoliated Deciduous Teeth

  • Akvilė Jarmalavičiūtė
  • Virginijus Tunaitis
  • Eglė Strainienė
  • Rūta Aldonytė
  • Arūnas Ramanavičius
  • Algirdas Venalis
  • Karl-Eric Magnusson
  • Augustas Pivoriūnas
Article

Abstract

Stem cells isolated from human adult tissues represent a promising source for neural differentiation studies in vitro. We have isolated and characterized stem cells from human exfoliated deciduous teeth (SHEDs). These originate from the neural crest and therefore particularly suitable for induction of neural differentiation. We here established a novel three-stage protocol for neural differentiation of SHEDs cells. After adaptation to a serum-free and neurogenic environment, SHEDs were induced to differentiate. This resulted in the formation of stellate or bipolar round-shaped neuron-like cells with subpopulations expressing markers of sensory neurons (Brn3a, peripherin) and glia (myelin basic protein). Commercial PCR array analyses addressed the expression profiles of genes related to neurogenesis and cAMP/calcium signalling. We found distinct evidence for the upregulation of genes regulating the specification of sensory (MAF), sympathetic (midkine, pleitrophin) and dopaminergic (tyrosine hydroxylase, Nurr1) neurons and the differentiation and support of myelinating and non-myelinating Schwann cells (Krox24, Krox20, apolipoprotein E). Moreover, for genes controlling major developmental signalling pathways, there was upregulation of BMP (TGF β-3, BMP2) and Notch (Notch 2, DLL1, HES1, HEY1, HEY2) in the differentiating SHEDs. SHEDs treated according to our new differentiation protocol gave rise to mixed neuronal/glial cell cultures, which opens new possibilities for in vitro studies of neuronal and glial specification and broadens the potential for the employment of such cells in experimental models and future treatment strategies.

Keywords

Neural differentiation Glial differentiation Neural crest Mesenchymal stem cells SHED 

Notes

Acknowledgments

This research was supported by a grant (No. MIP-084/2011) from the Research Council of Lithuania and the Swedish Research Council (KEM; No. 2010-3045). We would like to thank Dr. Arūnas Stirkė for technical assistance with the confocal microscopy.

Supplementary material

Supplementary video

After the induction of neural differentiation, SHEDs were monitored continuously in real time for approximately 120 h using Cell-IQ® PC Cell Imaging & Analysis System (CM Technologies) (WMV 23325 kb)

12031_2013_46_MOESM2_ESM.doc (246 kb)
Table S1 Differential expression of genes important for neurogenesis and neural stem cells during neural differentiation of SHEDs. We used RT2 Profiler™ PCR Arrays from SABiosciences, A QIAGEN company (Neurogenesis and Neural Stem Cell array, PAHS-404A). Gene expression levels were analysed using RT2 Profiler PCR Array Data Analysis software (version 3.5, QIAGEN) (DOC 246 kb)
12031_2013_46_MOESM3_ESM.doc (157 kb)
Table S2 Differential expression of genes important for cAMP/Ca2+ signalling during neural differentiation of SHEDs. We used RT2 Profiler™ PCR Arrays from SABiosciences, A QIAGEN company (Human cAMP/Ca2+ PathwayFinder array, PAHS-066A). Gene expression levels were analysed using RT2 Profiler PCR Array Data Analysis software (version 3.5, QIAGEN) (DOC 157 kb)
12031_2013_46_MOESM4_ESM.doc (173 kb)
Fig. S1 Differential expression of genes important for neurogenesis and neural stem cells during neural differentiation of SHEDs. Heatmap was generated using RT2 Profiler PCR Array Data Analysis software (version 3.5, QIAGEN) (DOC 173 kb)
12031_2013_46_MOESM5_ESM.doc (136 kb)
Fig. S2 Differential expression of genes important for cAMP/Ca2+ signalling during neural differentiation of SHEDs. Heatmap was generated using RT2 Profiler PCR Array Data Analysis software (version 3.5, QIAGEN) (DOC 136 kb)
12031_2013_46_MOESM6_ESM.jpg (896 kb)
Fig. S3 Formation of compact multilayer structures similar to the ganglions in differentiating SHEDs (JPEG 895 kb)

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Akvilė Jarmalavičiūtė
    • 1
    • 2
  • Virginijus Tunaitis
    • 1
  • Eglė Strainienė
    • 3
  • Rūta Aldonytė
    • 1
  • Arūnas Ramanavičius
    • 4
  • Algirdas Venalis
    • 1
  • Karl-Eric Magnusson
    • 5
  • Augustas Pivoriūnas
    • 1
  1. 1.Department of Stem Cell BiologyState Research Institute Centre for Innovative MedicineVilniusLithuania
  2. 2.Department of Neurobiology and Biophysics, Faculty of Natural SciencesVilnius UniversityVilniusLithuania
  3. 3.Department of Chemistry and BioengineeringVilnius Gediminas Technical UniversityVilniusLithuania
  4. 4.Department of Physical Chemistry, Faculty of ChemistryVilnius UniversityVilniusLithuania
  5. 5.Department of Clinical and Experimental MedicineLinköping UniversityLinköpingSweden

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