The process of regeneration developed by Fortuna Fix is based on the Axolotl Salamander’s well documented ability to regenerate.
Following injury, the surface of the wound is covered by a “wound epidermis” which is formed from epithelial cells that have migrated to the site of injury. Plasma, muscle fragments, cellular debris and lymphocytes accumulate under the wound epidermis. Dedifferentiated cells are released from the underlying tissue and accumulate at the site of injury, forming a blastema. The “dedifferentiated” cells within the blastema have re-accessed their own respective “cell development” programs for tissue formation, by undergoing reprogramming events to their own respective multipotent state, as if each somatic cell type had a memory of its origin which was triggered by the injury. Regeneration progresses and the multipotent cells divide, migrate and eventually differentiate down their own respective lineage, yielding within 40-50 days a perfectly restored limb or organ, in full respect of the original architecture and with no scar formation.
Fortuna Fix has perfected the same method for direct somatic cell reprogramming into neural stem (or precursor) cells without involving the pluripotent stage. The process is accomplished in vitro within 6-12 days by transient expression of reprogramming factors delivered using a synthetic plasmid (with no use of animal components or viruses and no integration into the genome).
One key feature of this process involves chromatin remodeling, by which the DNA wrapped within nucleosomes becomes accessible to transcription factors and the replication machinery. One of the reprogramming factors releases the chromatin and exposes the cell’s DNA to the other reprogramming factors that drive the reprogramming of the somatic cell to the neuronal stem cell by directly triggering the expression of at least one master or regulator gene resulting in the expression of a number of secondary genes characteristic of functional (and phenotypical) neural stem cells. Stable expression of the master and secondary genes is achieved by allowing the chromatin to remodel and lock into the neural stem cell epigenetic state. Following such a reprogramming event in vitro, the new cells are incubated in reagents and media components chemically optimized for neural stem cells. The resulting directly reprogrammed neural precursor cells (drNPCs) have the ability to continue proliferating for over 30 passages and can differentiate into a neural/glial/oligodendrocyte progenitor cell, ultimately differentiating to a neuron, an astrocyte or an oligodendrocyte. One of the effects of the direct cell reprogramming is the stabilization of the telomeres of the multipotent neural stem cells and the transient expression of telomerase, a clear sign of stemness and with consequent rejuvenation of the reprogrammed cells.
The process of direct cell reprogramming is now fully automated for high throughput manufacturing and can produce up to 100M cells per patient in approx 6 weeks.