Engraftment of mast cell-deficient mice with bone marrow derived-cultured mast cells (BMCMC) (PDF)

Mast cell donor mice: Ideally are around 6 weeks of age (4-12 weeks ok) and female

Mast cell recipient mice: Ideally around 5-6 weeks of age and of same genetic background, or as appropriate

1. Culture sufficient number of BMCMC for the mice you wish to engraft. Number of BMCMC required depends on the route of transfer; the following are guidelines, but should be refined for each experiment.

	Transfer Routes (per mouse):
	Intradermal:	1 x 106 cells in 20-50 µl injection “spots”; for example, to cover a 4 x 4 cm area of skin, use ~4 x 106 cells in a total of 200 µl, injected over the entire area
	Intraperitoneal:	2-4 x 106 cells in 200 µl
	Intravenous:	5-10 x 106 cells in 200 µl

2. Once BMCMC have been assayed for sufficient differentiation, ideally feed cells with fresh medium (may be supplemented with extra IL-3) one or two days before injection.

3. Count cells and resuspend in either plain cold DMEM or PBS. Keep cells on ice until ready for injection. Cells may be injected with a 30 gauge needle as required.

4. Time for engraftment depends on application and route of injection. Some general guidelines are as follows, though time needed for engraftment should be assessed for each experiment.

Intradermal: 4 – 8 weeks
Intraperitoneal: 6 – 8 weeks
Intravenous: 8 – 12 weeks