For cell transplantation: Difference between revisions
Jump to navigation
Jump to search
(Continuing directly from Liberase dissociation or from FACS)
File:1.jpg
File:2.jpg
| Line 13: | Line 13: | ||
*Follow the following protocol from here. | *Follow the following protocol from here. | ||
[[File:1.jpg]] | <li>[[File:1.jpg|200px]]</li> | ||
[[File:2.jpg]] | <li>[[File:2.jpg]]</li> | ||
[[File:Screenshot 2022-07-19 at 01.18.01.png]] | <li>[[File:Screenshot 2022-07-19 at 01.18.01.png]]</li> | ||
Revision as of 23:24, 18 July 2022
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.
