Isolation of Gingival Immune Cell Network from Murine Teeth Blocks

In This Article

Abstract

Source: Dutzan, N. et al., Isolation, Characterization and Functional Examination of the Gingival Immune Cell Network. J. Vis. Exp. (2016)

This video demonstrates the isolation procedure of the gingival immune cell network from a mouse model. The enzymatic digestion releases the gingival tissue from the attached teeth and bone, which is further mechanically digested, releasing immune cells from the gingiva. The isolated cells are ready for immunological experiments.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Prepare in Advance

Prepare Complete media: RPMI supplemented with 2 mM L-glutamine, 100 units/ml penicillin, 100 ug/ml streptomycin, and 10% FBS.
Prepare DNase media: 50 ml of RPMI media supplemented with 7.5 µg DNase (Make fresh, keep on ice at all times).
Prepare Collagenase-DNase media: 5 ml of DNase media plus 3.2 mg/ml of Collagenase Type IV (Make fresh, keep on ice at all times).
Pre-cool centrifuge to 4 °C.
War....

Access restricted. Please log in or start a trial to view this content.

Results

Tooth extraction, cross-section diagram, and histological slides at 4x and 10x magnification.
Figure 1: Isolation of murine gingiva. Illustration demonstrating the borders of dissection in the maxilla and the isolated gingival segment to be used for gingival tissue dissection. (B) Segment of the murine maxilla (molar area) with an outline for block dissection, and isolated block. Hema.......