Oral Delivery of Transformed Yeast Cells into the Mouse Intestinal Immune System

Published: May 31, 2024

Abstract

Source: Hudson, L. E. et al., Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice. J. Vis. Exp. (2016)

This video showcases the oral administration of transformed yeast cells into a mouse model. The transformed yeast cells are orally administered using a gavage technique, reaching the mouse's stomach. Later, the cells migrate to the small intestine, where microfold cells engulf them and facilitate their transfer to Peyer's patches and gain access to the intestinal immune system.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. UV Mutagenesis to Generate Auxotrophic Yeast Strains Generate survival curves to determine needed doses of ultraviolet (UV) irradiation Prepare YPD (yeast extract peptone dextrose) media and other reagents listed in Table 1 according to standard procedures and inoculate single…

Representative Results

Figure 1. Yeast colonies grown on YPD media. Example YPD plate showing viable colony forming units (CFU) of probiotic yeast after UV irradiation. Cells were serially diluted such that individual CFU can be distinguished and counted. Figure 2. Survival c…

Offenlegungen

The authors have nothing to disclose.

Materials

SmartSpec 3000 Spectrophotometer BioRad 170-2501 Example of spectrophotometer for determining cell concentration and OD600 of yeast cultures
New Brunswick Roller Drum Eppendorf M1053-4004 Example of roller drum for yeast culture incubation
UV Stratalinker 2400 Stratagene 400075-03 Example stratalinker
Stuart Colony Counter SC6PLUS 11983044 Fisher Scientific Plate stand with magnification records colony count upon sensing pressure from pen
Scienceware Colony Counter F378620002 Bel-Art Scienceware Hand held colony counter pen
Replica plating device Fisherbrand 09-718-1 Example of replica plating stand and pads
Velveteen squares Fisherbrand 09-718-2
L shaped sterile cell spreaders Fisherbrand 14665230
Deoxyribonucleic acid, single stranded from salmon testes Sigma-Aldrich D7656-1ML Example carrier DNA for yeast LiOAc transformation
Gavage needles Braintree Scientific N-PK 002 For mice 15-20 g, the suggested needle is a 22 gauge (1.25 mm ball), 1 in long, straight reusable gavage needle. For mice weighing greater than 20 g, 20 gauge or larger straight or curved gavage needles may be used
1mL sterile slip-tip disposable tuberculin syringe Becton Dickinson BD 309659
Blunt forceps such as Electron Microscopy Sciences 7" (178 mm) serrated tip, broad grip forceps Electron Microscopy Sciences 77937-28 Example of blunt forceps needed for dissection
Straight and curved dissection scissors Electron Microscopy Sciences 72966-02 and 72966-03 Examples of scissors needed for dissection

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Oral Delivery of Transformed Yeast Cells into the Mouse Intestinal Immune System. J. Vis. Exp. (Pending Publication), e22271, doi: (2024).

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