MSC Seeded Scaffold Preparation: A Technique to Prepare Scaffold Seeded with Mesenchymal Stem Cells for Surgical Implantation into Murine Model
Transcrição
Cancer cells produce chemotactic homing factors to attract the mesenchymal stem cells or MSCs from the blood vessels towards the tumor location.
To study the effect of these cells on tumor growth in vitro, begin by taking a sterilized scaffold substrate. Transfer it to a well-plate containing desired media to pre-wet the scaffold. Place it on a dry surface to remove excess media sticking to the scaffold for better cell adhesion. Transfer the partially dried scaffold to a fresh well. Seed a drop of MSC suspension onto the scaffold.
Additionally, add more media in the corners of the well to create a hydration chamber that prevents media evaporation from the drop during subsequent incubation. Incubate to allow the MSCs to adhere to the surface of the scaffold. Flip over the scaffold to expose the non-seeded surface. Pipette a fresh drop of MSC suspension on this side and let MSCs adhere.
After MSCs have adhered to both sides of the scaffold, cover it with more media ensuring that all the cells are in contact with the media. Incubate till the MSCs grow and infiltrate within the interstices and pores of the scaffold. The MSC seeded scaffold is ready for surgical implantation.
