A novel stem cell for auditory neuron regeneration.

Transcription

1 A novel stem cell for auditory neuron regeneration. M.A.Huisman*, R. el Seady*, C.F.H.M.Löwik # en J.H.M.Frijns* Departments of Ear, Nose & Throat* and Molecular Imaging #, Leiden University Medical Centre, The Netherlands

2 The problem In mammals: No regeneration of cochlear sensoric epithelium and the associated neural components

3 Cochlear implant Greatly benefits patients In trial is improvement of: hearing with background noise music perception Hair cells per patient effectivity CI Auditory neuron regeneration Auditory neurons

4 The choice of experimental therapeutics: Neurotrophic factors (NF) Stem cells

5 eurotrophic Factors Brain-derived neurotrophic factor (BDNF) Neurotrophin-3 (NT-3) Nerve Growth Factor (NGF) Fibroblast derived growth factor (FGF) Glial cell-derived neurotrophic factor (GDNF) NF mini osmotic pump and reservoir...on location by genetic manipulated cells...on location by nano particles...+ chronic electrical stimulation(ces)

6 So far: GF therapy for auditory neurons It works, in vitro and in vivo, CES increases effect Pettingill LN Richardson RT. 2008; Okano Achterberg M Weissleder R Dobson J.2008 Praetorius M.2007

7 Stem cell therapy: embryonic & postnatal stem cells

8 Stem cell potency pluripotent multipotent multipotent multipotent

9 Stem cell harvesting: embryonic stem cells From the inner cell mass of the blastocyst ( cell stage) High proliferation and differentiation potential, migrative Courtesy: Molecularstation.com but: Ethical problems, tumor prone Groups of hundreds embryonic miuse stem cells on feeder cells. S.Heller

10 Stem cell harvesting: postnatal stem cells + Bone marrow high yield Umbilical cord Neural crest Inner ear high migration IESC Nose Hair follicle autologue transfusion, easy harvesting

11 Stem cell therapy: postnatal stem cells Less potential than ESC: Proliferation, differentiation, potency But: No tumor formation Some postnatal stem cells show pluripotency

12 So far: stem cell therapy for auditory neurons In vivo: Survival, migration and differentiation IESC ESC ansc/ncsc BMSC OSC Neural progenitors in the cochlea of the gerbil (after Ouabain treatment) Fritzsch B Li H Oshima K Corrales CE 2006 Sekiya 2007

13 What Which Why? happens stem therapy? cell? in Leiden? not tumor prone Hair no ethical follicle problem Stem stem cells cells autologue transfusion January 2007 multipotent neural lineage pluripotency easy harvesting same origin as inner ear cells

14 Which Why? stem therapy? cell? not tumor prone Hair no ethical follicle problem Stem stem cells cells High potential for both, repopulation and regrowth of SGN autologue transfusion multipotent neural lineage pluripotency Promising results in the USA and Japan. easy harvesting same origin as inner ear cells

15 Which Why? stem cell? not tumor prone Hair no ethical follicle problem stem cells High potential for both, regrowth and repopulation of SGN autologue transfusion multipotent neural lineage pluripotency Promising results in the USA and Japan. easy harvesting same origin as inner ear cells

16 Which Why? stem cell? not tumor prone Hair no ethical follicle problem stem cells autologue transfusion multipotent neural lineage pluripotency easy harvesting same origin as inner ear cells

17 Why? not tumor prone no ethical problem autologue transfusion multipotent neural lineage pluripotency easy harvesting same origin as inner ear cells

18 The question Are hair follicle stem cells potential candidates for auditory neuron regeneration? Sufficient stem cells Neurons with SGN characteristics Myelinating glia cells

19 Harvesting hair follicle stem cells (EPI-NCSC) Poster El Seady

20 EPI- CSC from different species Mouse, chinchilla, guinea pig 1 week 1.5 weeks 90% positivity for neural stem cell marker nestin Human >95% nestin positive

21 EPI- CSC yield: Sufficient stem cells? Average yield per follicle 2x10 5 cells 3 hair follicles are sufficient for transplantation

22 eurons from EPI- CSC 70% neurons βiii-tubulin-positive Migrative β-tubulin Network formation Neurofilaments 100 um

23 Cells from EPI- CSC-neurons Neurons with SGN characteristics Bipolarity 60 % bipolar TrkB/C coexpression BDNF/ NT3 growth factor receptors 100 um

24 Cells from EPI- CSC Myelinating glia cells Schwann cell 20% Schwann cells 30% Oligodendrocytes Distinct morphology Oligodendrocyte 5 um Myelinating cells 10 um

25 The question Are hair follicle stem cells potential candidates for auditory neuron regeneration? Sufficient stem cells Neurons with SGN characteristics Myelinating glia cells

26 The question Are hair follicle stem cells potential candidates for auditory neuron regeneration? YES!

27 Future perspectives Detection in the living animal poster El Seady Functionality, also in electrical field(ci) Xenotransplantation, animal model autologue transplantations in humans Autologue transplantation, animal model

28 References Growth factor therapy Pettingill LN. et al. Review IEEE Trans Biomed Eng Jun;54(6 Pt 1): Richardson RT. et al. Expert Opin Drug Deliv. 2008; Oct;5(10): Okano et al.mol Ther. 2006; 14: Achterberg M. et al. Hearing Research. 2008;244:25-34 Weissleder R. et al. Nature Biotechnology. 2005; 23: Dobson J. Nature Nanotechnology March;3: Praetorius at al. Acta Otolaryngol.2007 May;127(5): Stem cell therapy inner ear: Fritzsch B. et al. Bioessays. Review 2006 Dec;28(12): Li H. et al. PNAS 2003, 11( 100) no. 23; Oshima K. et al. JARO 2007; 8:18-31 Corrales CE et al.j Neurobiol Nov;66(13): Sekiya et al. Eur J Neurosc 2007(25); Rask-Andersen H et al. Hearing Research;2005: Sieber Blum, Dev Dyn 2006 Detection Couillard-Despres et al Eur J Neurosci Sep;24(6): Aigner L., Kruttwig K., Löwik C., pers comm Uncomplicated differentiation into bipolar neurons and myelinating glia. El Seady R., Huisman MA.,Lowik CW., Frijns JH. Biochem Biophys Res Commun Nov 14;376(2):