One of the challenges in clinical translation of cell-replacement therapies is the definition of optimal cell generation and storage/recovery protocols which would permit a rapid preparation of cell-treatment products for patient administration. Besides, the availability of injection devices that are simple to use is critical for potential future dissemination of any spinally targeted cell-replacement therapy into general medical practice. Here, we compared the engraftment properties of established human-induced pluripotent stem cells (hiPSCs)-derived neural precursor cell (NPCs) line once cells were harvested fresh from the cell culture or previously frozen and then grafted into striata or spinal cord of the immunodeficient rat. A newly developed human spinal injection device equipped with a spinal cord pulsation-cancelation magnetic needle was also tested for its safety in an adult immunosuppressed pig. Previously frozen NPCs showed similar post-grafting survival and differentiation profile as was seen for freshly harvested cells. Testing of human injection device showed acceptable safety with no detectable surgical procedure or spinal NPCs injection-related side effects.

• Keywords
• cryopreservation, human induced pluripotent stem cells (hiPSCs), human injection device, immunosuppressed adult pig, neural precursor cells (NPCs), spinal cord,
• MeSH
• Cell Differentiation physiology   MeSH
• Adult MeSH
• Genetic Vectors genetics   MeSH
• Induced Pluripotent Stem Cells * physiology   transplantation   MeSH
• Rats MeSH
• Humans MeSH
• Spinal Cord MeSH
• Brain MeSH
• Neural Stem Cells * physiology   transplantation   MeSH
• Specimen Handling methods   MeSH
• Tissue and Organ Harvesting methods   MeSH
• Swine MeSH
• Cellular Reprogramming * genetics   physiology   MeSH
• Graft Survival physiology   MeSH
• Injections, Spinal * adverse effects   instrumentation   methods   MeSH
• Stem Cell Transplantation * adverse effects   instrumentation   methods   MeSH
• Sendai virus MeSH
• Treatment Outcome MeSH
• Animals MeSH
• Check Tag
• Adult MeSH
• Rats MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

A promising therapeutic strategy for amyotrophic lateral sclerosis (ALS) treatment is stem cell therapy. Neural progenitors derived from induced pluripotent cells (NP-iPS) might rescue or replace dying motoneurons (MNs). However, the mechanisms responsible for the beneficial effect are not fully understood. The aim here was to investigate the mechanism by studying the effect of intraspinally injected NP-iPS into asymptomatic and early symptomatic superoxide dismutase (SOD)1G93A transgenic rats. Prior to transplantation, NP-iPS were characterized in vitro for their ability to differentiate into a neuronal phenotype. Motor functions were tested in all animals, and the tissue was analyzed by immunohistochemistry, qPCR, and Western blot. NP-iPS transplantation significantly preserved MNs, slowed disease progression, and extended the survival of all treated animals. The dysregulation of spinal chondroitin sulfate proteoglycans was observed in SOD1G93A rats at the terminal stage. NP-iPS application led to normalized host genes expression (versican, has-1, tenascin-R, ngf, igf-1, bdnf, bax, bcl-2, and casp-3) and the protection of perineuronal nets around the preserved MNs. In the host spinal cord, transplanted cells remained as progenitors, many in contact with MNs, but they did not differentiate. The findings suggest that NP-iPS demonstrate neuroprotective properties by regulating local gene expression and regulate plasticity by modulating the central nervous system (CNS) extracellular matrix such as perineuronal nets (PNNs).

• Keywords
• ALS, iPS, motoneuron death, neurodegeneration, plasticity, proteoglycans, stem cells, transplantation,
• MeSH
• Amyotrophic Lateral Sclerosis therapy   MeSH
• Induced Pluripotent Stem Cells cytology   MeSH
• Rats MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Neural Stem Cells cytology   metabolism   transplantation   MeSH
• Neuronal Plasticity * MeSH
• Nerve Growth Factors genetics   metabolism   MeSH
• Peripheral Nerves physiology   MeSH
• Rats, Sprague-Dawley MeSH
• Apoptosis Regulatory Proteins genetics   metabolism   MeSH
• Nerve Regeneration MeSH
• Tenascin genetics   metabolism   MeSH
• Stem Cell Transplantation methods   MeSH
• Versicans genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Nerve Growth Factors MeSH
• Apoptosis Regulatory Proteins MeSH
• Tenascin MeSH
• Versicans MeSH