Isolating Induced Neural Progenitor Cell Colonies Derived From Adult Human Fibroblasts
Transcription
Begin with an adherent culture of induced neural progenitor cell colonies derived from adult human fibroblasts in a laminin-coated multi-well plate.
Wash the colonies with a buffer to remove non-adherent cells, then add fresh buffer.
Scrape around individual colonies to remove surrounding cells.
Transfer the colonies to fresh laminin-coated wells containing neuroinduction medium.
Mechanically disaggregate the colonies to obtain a single-cell suspension.
Laminin, an extracellular matrix protein, interacts with the surface proteins of the progenitors, facilitating cell adhesion to the well surface.
Add a small molecule inhibitor that enters the cells and blocks specific protein kinases, preventing cellular apoptosis.
Growth factors and small molecules present in the neuroinduction medium facilitate the proliferation of the progenitors.
Periodically replace the medium to replenish nutrients, facilitating continued progenitor proliferation.
A confluent culture, or a fully covered cell layer, indicates the successful expansion of induced neural progenitor cells.
