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…
Divulgaciones
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).