New possible canine testis tissue extraction and culture
Faculty Mentor
Justin Bastow
Presentation Type
Oral Presentation
Start Date
5-7-2024 10:45 AM
End Date
5-7-2024 11:05 AM
Location
PAT 328
Primary Discipline of Presentation
Biology
Abstract
A novel method of obtaining live thin sections of dog testes was developed. In contrast with traditional cell culture system, where tissues are disrupted into single cells with proteolytic enzymes and cultivated in optimal growth media, dog testes thin sections were generated with an electric slicer (the type that can be found at any supermarket). Thus, the experiment attempted to culture mammalian organ sections while being cost-effective. If successful, this system would enable mammalian organotypic cell cultures to be widely employed in reproductive biology. Dog testes were obtained from a local spay/neuter veterinary clinic and sliced into 400-600µm thickness with the electric slicer. The sections were cultured in Petri dishes containing Tissue Culture Media-199 (TCM-199). The sections media was equilibrated with a blood gas mixture (7% CO2 :7% O2 : balanced N2) and cultured in an air-tight plastic chamber at 37 degrees Celsius in a water-jacketed incubator. As time elapsed, the sections were observed to thicken, curve on edges and transform into tiny testis-like structures. These changes occurred after 7 to 10 days of culture and progressed until staining commenced. The sections were confirmed to be alive after 21 days of culture by live/dead cell staining. Through this preliminary work, future studies can employ this system to potentially obtain viable mature sperm cells in vitro.
Recommended Citation
Becerra, Jonathan; Singh, Ramanpreet; Merritt, Sadie; Nguyen, Nguyen K.; Sanborn, Michael; and Sundheim, Taiyo, "New possible canine testis tissue extraction and culture" (2024). 2024 Symposium. 5.
https://dc.ewu.edu/srcw_2024/op_2024/o4_2024/5
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
New possible canine testis tissue extraction and culture
PAT 328
A novel method of obtaining live thin sections of dog testes was developed. In contrast with traditional cell culture system, where tissues are disrupted into single cells with proteolytic enzymes and cultivated in optimal growth media, dog testes thin sections were generated with an electric slicer (the type that can be found at any supermarket). Thus, the experiment attempted to culture mammalian organ sections while being cost-effective. If successful, this system would enable mammalian organotypic cell cultures to be widely employed in reproductive biology. Dog testes were obtained from a local spay/neuter veterinary clinic and sliced into 400-600µm thickness with the electric slicer. The sections were cultured in Petri dishes containing Tissue Culture Media-199 (TCM-199). The sections media was equilibrated with a blood gas mixture (7% CO2 :7% O2 : balanced N2) and cultured in an air-tight plastic chamber at 37 degrees Celsius in a water-jacketed incubator. As time elapsed, the sections were observed to thicken, curve on edges and transform into tiny testis-like structures. These changes occurred after 7 to 10 days of culture and progressed until staining commenced. The sections were confirmed to be alive after 21 days of culture by live/dead cell staining. Through this preliminary work, future studies can employ this system to potentially obtain viable mature sperm cells in vitro.
