For cell transplantation
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(Continuing directly from Liberase dissociation or from FACS)
Cell transplantation
- 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.
