Cell transplantation

(Continuing directly from Liberase dissociation or from FACS)

Resuspend the cell pellet with 100-200uL AMEM(0) depending on the size of the pellet.
Count the cells using a hemocytometer.
- Standard protocol: 5uL cell solution + 5uL trypan blue, count the cells in 5 blocks

cell number/5 x 20 = cell number/uL

Calculate the required volume (<200uL) for transplantation: 10k cells per transplant.
- For example, minimum 50k donor cells are required for 5 transplants.
- If the required volume is larger than 200uL, spin down and resuspend the cells with a smaller volume.
Load the required volume of the donor cell solution into a sealed yellow tip.
Centrifuge the sealed yellow tips in a 50mL falcon tube. 300xrcf, 20 degree, 5 minutes.
Follow the following protocol from here.

For cell transplantation