Candida albicans Biofilm Formation in an In Vivo Mouse Model
Published: November 30, 2023
Abstract
Source: Kucharíková, S., et al. Candida albicans Biofilm Development on Medically-relevant Foreign Bodies in a Mouse Subcutaneous Model Followed by Bioluminescence Imaging. J. Vis. Exp. (2015).
This video demonstrates the development of an in vivo mouse model to study the biofilm formation of Candida albicans in subcutaneous tissue. The experiment involves implanting Candida-infected devices in the animal's back region for biofilm development.
Protocol
1. C. albicans Growth Twenty-four hours before the initiation of the animal experiment, prepare yeast extract peptone dextrose, YPD plates by adding 10 g of yeast extract granulated, 20 g of bacteriological peptone, and 15 g of granulated agar. Makeup volume to 900 ml with Milli-Q water and autoclave. Add 50 ml of sterile 40% glucose. Mix thoroughly and pour into Petri dishes. Leave agar plates to cool and solidify. NOTE: In this study, use two strai…
Representative Results
Figure 1: Major steps during the animal surgery. (A) The procedure of catheter implant. (1) Place the anesthetized animal on a warm pad containing paper tissue and apply the ophthalmic ointment to the eyes. (2) Shave the lower part of the back and disinfect. (3) Create two small (approx. 0.5 cm) incisions through the skin on the left and on the right side of the back. Create a subcutaneous tunne…
Açıklamalar
The authors have nothing to disclose.
Materials
| Yeast extract granulated | Merck | MERC1.03753.0500 | |
| Bacteriological peptone | Oxoid | LP037B | |
| Agar granulated | Difco | 214530 | |
| Phosphate buffered saline | Prepared in the laboratory | for 1L of 10x PBS: 80 g NaCl, 2 g KCl, 14.4 g Na2HPO4, 2.4 g KH2PO4 | |
| RPMI1640 with L-glutamine and without sodium carbonate | Sigma | R6504-1L | Prepare according the protocol for Candida albicans drug susceptibility testing |
| fetal bovine serum (FBS) | Sigma | F7524 | |
| Polyurethane tripe-lumen intravenous catheter piece (2.4 mm diameter, Certofix Trio S730) | BBraun | CV-15703 | CV-15703 |
| Dexamethasone | Fagron SAS, France | 611139 | Immunosuppressant (stock solution 10 mg/ml) |
| Insulin syringes (0.3 ml) | Terumo Myjector 29G | 324826 | For injection of coelenterazine |
| Electric razor | For small animals | ||
| Sterile surgical tools | Scissors, 2 pairs of tweezers, scalpel | ||
| Heating pad | Leica | 14042321474 | |
| Skin suture | Johnson&Johnson | K890H | Surgical thread, needle |
| BLI camera (IVIS Spectrum) | Perkin Elmer, Alameda | IVISSPE | |
| Living Image software | Perkin Elmer, Alameda | (version 4.2) |
Bu Makaleden Alıntı Yapın
Candida albicans Biofilm Formation in an In Vivo Mouse Model. J. Vis. Exp. (Pending Publication), e21797, doi: (2023).