Condition optimization for electroporation transfection in horse skeletal muscle satellite cells.
Abstract: Satellite cells are an important cellular model for studying muscle growth and development and mammalian locomotion-related molecular mechanisms. In this study, we investigated the effects of voltage, pulse duration, and DNA dosage on horse skeletal muscle satellite cells' electroporation transfection efficiency using the eukaryotic expression plasmid Td Tomato-C1 (5.5 kb) encoding the red fluorescent protein gene mainly based on fluorescence-positive cell rate and cell survival rate. By comparison of different voltages, pulse durations, and DNA doses, horse skeletal muscle satellite cells have nearly 80% transfection efficiency under the condition of voltage 120 V, DNA dosage 7 µg/ml, and pulse duration 30 ms. This optimized electroporation condition would facilitate the application of horse skeletal muscle satellite cells in genetic studies of muscle function and related diseases.
Publication Date: 2023-11-20 PubMed ID: 37982395DOI: 10.1080/10495398.2023.2280664Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article focuses on identifying the optimal conditions for electroporation transfection, a method for introducing foreign DNA into cells, in horse skeletal muscle satellite cells.
Understanding the Study
- The researchers aimed to optimize the conditions for successfully introducing foreign DNA into horse skeletal muscle satellite cells.
- These cells are an important model for studying muscle growth, development, and mammalian locomotion-related molecular mechanisms.
- The efficiency and success rate of the electroporation transfection process were studied under varying parameters like voltage, pulse duration, and DNA dosage.
- The foreign DNA used in the study was encoded using the eukaryotic expression plasmid Td Tomato-C1 (5.5 kb), harboring the red fluorescent protein gene. The success rate of the transfected cells was gauged by checking the number of cells emitting red fluorescence.
Findings of the Study
- It was found that the high transfection efficiency of close to 80% was achieved under specific conditions:
- The transfection process was optimized under a voltage setting of 120V.
- The pulse duration was optimized at 30ms.
- A DNA dosage of 7 µg/ml was found to be optimum.
Significance of the Research
- This research contributes to a better understanding of the optimal conditions under which the electroporation transfection process can be carried out efficiently in horse skeletal muscle satellite cells.
- It thus aids in setting a standardized procedure for the electroporation transfection in these cells, reducing experimental errors and inconsistencies and enhancing research outcomes.
- Such optimization should also help facilitate the application of horse skeletal muscle satellite cells to genetic studies related to muscle function and relevant diseases, thereby boosting the potential of such research in both academic and therapeutic contexts.
Cite This Article
APA
Ulaangerel T, Yi M, Budsuren U, Shen Y, Ren H, Demuul B, Bai D, Dorjgotov D, Davaakhuu G, Jambal T, Dugarjav M, Bou G.
(2023).
Condition optimization for electroporation transfection in horse skeletal muscle satellite cells.
Anim Biotechnol, 1-9.
https://doi.org/10.1080/10495398.2023.2280664 Publication
Researcher Affiliations
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- School of Animal Science and Biotechnology, Mongolian University of Life Sciences, Ulaanbaatar, Mongolia.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- School of Industrial Technology, Mongolian University of Science and Technology, Ulaanbaatar, Mongolia.
- Institute of General and Experimental Biology, Mongolian Academy of Science, Ulaanbaatar, Mongolia.
- School of Industrial Technology, Mongolian University of Science and Technology, Ulaanbaatar, Mongolia.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
- lnner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
