Difference between revisions of "For cell transplantation"

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*Follow the following protocol from here.
 
*Follow the following protocol from here.
  
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Revision as of 23:37, 18 July 2022

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.
  • Screenshot 2022-07-19 at 01.18.01.png
  • 2.png