Ajankohtaista silmän kantasolututkimuksesta

Transcription

1 Ajankohtaista silmän kantasolututkimuksesta Tanja Ilmarinen FT, vanhempi tutkija No commercial relationships

2 Kantasolututkimusta Tampereella Tampereen yliopisto - Lääketieteen ja biotieteiden tiedekunta Solu- ja kudosteknologia Heli Skottman - Silmäryhmä Katriina Aalto-Setälä - Fysiologia, Sydänryhmä Susanna Miettinen - Aikuisten kantasolujen tutkimusryhmä Susanna Narkilahti - Neuroryhmä Timo Ylikomi Solubiologia Tero Järvinen - Kudosregeneraatio BioMediTech instituutti - Tampereen yliopiston ja Tampereen teknillisen yliopiston yhteinen instituutti

3 Human pluripotent stem cells Human embryonic stem cells (hescs) Target cells HLA-matched cell banks Human induced pluripotent stem cells (hipscs) Modified from Carr et al, 2013

4 Stem cell applications AMD

5 Cell-based therapy for retinal degeneration Li LX, Turner JE. Inherited retinal dystrophy in the RCS rat: prevention of photoreceptor degeneration by pigment epithelial cell transplantation. Exp Eye Res Macular translocation high rate of complications Autologous RPE-Choroid Patch graft high rate of complications Subretinal injection of a suspension of RPE cells promising approach cell source?

6 RPE from pluripotent stem cells

7 Clinical compatibility what is needed? Clinically compatible materials and methods Avoiding xeno-products and highly variable components such as serum and feeder cells Efficient production of large homogeneous cell batches Production of cells which are terminally differentiated, functionally compentent and robust (can endure additional transplantation induced cellular stress) /articleno/18757 Strauss, 2005 Joddar et al 2014 Biomater. Sci., 2014, 2,

8 Our approach hpsc-rpe : Added safety by xeno and feeder-free produced cells d1 d5 Week 4 Week 10 Week 19 Hongisto et al. submitted Clinically compatible, easy transition to GMP production Two stage differentiation strategy: initial neuroectodermal induction with WNT and TGF-beta inhibitors in suspension followed by adherent differentiation on optimised ECM After differentiation 9-11 week maturation culture on inserts to achieve apical-basal polarity and functional competence Xeno-free cryobanking and recovery of RPE

9 Our approach Functional and robust RPE from hpscs Production of cells which are terminally differentiated, functionally competent and robust Day 35 End point End point Hongisto et al. submitted Viheriälä et al. In preparation

10 Our approach hpsc-rpe: Culture substrate affects cell functionality in vitro Viheriälä et al. In preparation

11 Our approach hpsc-rpe: Culture substrate affects cell functionality in vitro CIV ATP1 CIV+521 ATP1 Viheriälä et al. In preparation

12 Method of transplantation suspension or sheet? Attachment on diseased/aged Bruch s membrane vs. Invasiveness of surgery Risks from supportive substrate

13 Our approach hpsc-rpe: Safety (and functionality) in vivo Sheet transplantations in rabbits in collaboration with Prof. Hannu Uusitalo (UTA), MD Peeter Kööbi (Tampere University Hospital) and MD Boris Stanzel (University of Bonn) Polyimide scaffold without specialized instrument Polyester scaffold using a custom-made transplantation instrument Sheet transplantations in non-human primates in collaboration with MD Boris Stanzel and MD Gavin Tan (Singapore Eye Research Institute) Polyester scaffold using a custom-made transplantation instrument Removal of endogenous RPE Xeno- and feeder-free hesc-rpe 1 month follow-up, 14 animals Primary outcome measure: To assess submacular integration of hesc-rpe grafts in a fovea-containing large-eyed model with disrupted blood retinal barrier Safety Secondary outcome measure: Effect of surgical protocols on macular dynamics assessed by ioct

14 Safety (and functionality) in vivo: BioMediTech institute to Singapore National Eye Centre, Singapore Cell shipment before shipment TER Ω cm 2 after shipment Shipment at RT in medium optimized for short-term culture in ambient CO2 5 separate shipments Shipment times 32h -33h (54h) Map data 2017 Google, INEGI United States 1000 mi

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16 Safety (and functionality) in vivo: Transplantation

17 Safety (and functionality) in vivo: Preliminary results Structural and functional integration of submacular hesc- RPE xenografts in a novel monkey model with disrupted RPE barrier So far no cell-related safety issues such as tumor formation detected The current implantation technique requires further development

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19 Clinical trials with hpsc-rpe Ongoing in USA, Europe and Asia: hesc-rpe suspension hipsc/hesc-rpe sheet with or without an artificial scaffold Currently under evaluation: Safety and dose-response Efficacy Method of transplantation hesc vs ipsc

20 Private donations

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