Supplementary Materials

Supplementary Material for:

Survival of syngeneic and allogeneic iPSC–derived neural precursors after spinal grafting in minipigs

Jan Strnadel, Cassiano Carromeu, Cedric Bardy, Michael Navarro, Oleksandr Platoshyn, Andreas N. Glud, Silvia Marsala, Jozef Kafka, Atsushi Miyanohara, Tomohisa Kato Jr., Takahiro Tadokoro, Michael P. Hefferan, Kota Kamizato, Tetsuya Yoshizumi, Stefan Juhas, Jana Juhasova, Chak-Sum Ho, Taba Kheradmand, PeiXi Chen, Dasa Bohaciakova, Marian Hruska-Plochan, Andrew J. Todd, Shawn P. Driscoll, Thomas D. Glenn, Samuel L. Pfaff, Jiri Klima, Joseph Ciacci, Eric Curtis, Fred H. Gage, Jack Bui, Kazuhiko Yamada, Alysson R. Muotri, Martin Marsala*

*Corresponding author. Email: mmarsala{at}ucsd.edu

Published 9 May 2018, Sci. Transl. Med. 10, eaam6651 (2018)
DOI: 10.1126/scitranslmed.aam6651

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Porcine fibroblast-derived iPSCs generate ectoderm, mesoderm, and endoderm cell derivatives in vitro and in vivo.
  • Fig. S2. Previously frozen, in vitro–expanded porcine iPSC-NPCs show stable karyotype and generate neural derivatives (neurons, astrocytes, and oligodendrocytes) after in vitro induction.
  • Fig. S3. Long-term grafted (7 to 10 months) porcine iPSC-NPCs in rat striata show protein- and mRNA-defined signature that is consistent with mature porcine CNS tissue.
  • Fig. S4. Porcine iPSC-NPCs grafted into rat striata show no tumor formation and incomplete myelinization at 7 months after grafting.
  • Fig. S5. Porcine iPSC-NPC-EGFP+ grafts in rat striata show normal vascularization and no changes in tumor suppressors or proto-oncogenes at 7 to 10 months after grafting.
  • Fig. S6. iPSC-NPCs grafted into syngeneic pig spinal cord in the absence of immunosuppression show long-term survival and neuronal and glial differentiation at 3 months after transplantation.
  • Fig. S7. Porcine iPSC-NPCs grafted spinally in allogeneic, transiently immunosuppressed (1-month immunosuppression) pigs with previous spinal traumatic injury show neuronal and glial differentiation at 3.5 months after grafting.
  • Fig. S8. Spinally grafted iPSC-NPCs in allogeneic, spinally injured pig with transient immunosuppression (1 month) show extensive neuronal (NeuN) differentiation at 3.5 months after grafting.
  • Fig. S9. iPSC-NPCs grafted spinally in allogeneic pig with previous spinal injury do not form tumors and show incomplete myelination at 3.5 months after grafting.
  • Fig. S10. Reprogramming factors (OCT4 and KLF4) are silenced in mature iPSC-NPC grafts in rat striata or spinal cord in allogeneic pig with previous spinal traumatic injury.
  • Fig. S11. Long-term grafted iPSC-NPCs in rat striata or spinal cord of allogeneic pig show only occasional presence of Sendai virus–associated protein in grafted cells and show no change in expression of immunogenic genes.
  • Table S1. Quantitative analysis of neuronal and glial differentiation in EGFP grafts.
  • Table S2. Electrophysiological properties of three transplanted iPSC neurons into the striatum of a rat at 8 months after grafting.
  • Table S3. mRNA-sequencing species sorting quantification.
  • Table S4. SLA genotypes of the iPSC-NPC donor and the allogeneic graft recipients.
  • Table S5. Antibodies used for flow cytometry and immunofluorescence staining.

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Other Supplementary Material for this manuscript includes the following: