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.
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
2. Isolation, Dissection, and Cell Isolation from Gingival Blocks
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. Hematoxylin and Eosin stain (H&E) of a maxillary molar segment with gingival tissues outlined is shown in low (C) and higher magnification in (D). Original magnifications indicated.
The authors have nothing to disclose.
Fine Scissors | Fine science tools | 14058-11 | |
Scalpel Handle #3 | Fine science tools | 10003-12 | |
Scalpel Blades #10 | Fine science tools | 10010-00 | Sterile |
Splinter Forceps | Integra Miltex | 6-304 | |
Needles with regular bevel | BD Medical | 305109 | 27G, 12.7 mm length |
Monoject syringes | Covidien | 8881513934 | Luer-lock tip, 3mL |
PBS, pH 7.4 | Life Technologies | 10010-049 | Without Calcium and Magnesium |
RPMI 1640 | Lonza | 12-167F | Without L-glutamine |
DNase I from bovine pancreas | Sigma-Aldrich | DN25-1G | |
Collagenase type IV | Gibco (by Life technologies) | 17104-019 | |
Fetal Bovine Serum | Gemini Bio-products | 100-106 | |
Gentamicin 50 mg/ml | Quality biological | 120-098-661EA | |
Pen Strep | Gibco (by Life technologies) | 15140-122 | |
L-Glutamine | Gibco (by Life technologies) | 25030-081 | |
0.5M EDTA pH 8.0 | Quality biological | 351-027-721EA | |
50 mL tubes | Corning | 352070 | Polypropylene, sterile |
70 μM Cell Strainers | Corning | 352350 | |
Petri dishes | Corning | 351029 | Sterile |