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Calu-3/A-549 mixed cells as a replacement for primary rhesus monkey kidney cells for virus detection
Stacy Weber Fening1, Joseph A Jollick, Yung T Huang
1Case Western Reserve University, 2085 Adelbert Road, Cleveland, OH 44106, United States.
Background:
Primary kidney cells derived from rhesus macaques (pRhMK) are heavily relied upon for the detection and culture of a wide range of clinically relevant viruses. The use of these cultures is problematic due to the possible presence of endogenous viruses, the need to sacrifice a primate, and the inherent variance found in primary cultures.
Objective:
To develop a continuous cell line or mixed cell co-culture that could replace dependence on pRhMK cells.
Study Design:
Cells from the Calu-3 and A-549 cell lines were used to prepare mixed cell monolayers that were compared to pRhMK cells for their ability to detect respiratory viruses, measles, mumps, enteroviruses, and herpes viruses. Clinically derived and laboratory virus strains were used for these comparisons in culture plates or 16 mm tubes.
Results:
Calu-3/A-549 cells are more sensitive than pRhMK for the detection of adenovirus, enteroviruses and herpes simplex virus and are about equally sensitive for the detection of other respiratory viruses, measles, mumps and varicella-zoster virus.
Conclusions:
Calu-3/A-549 cells are an equivalent or better alternative to pRhMK cells for the detection of many clinically relevant viruses.
Insights
A new mixed cell culture using Calu-3 and A-549 cells offers a superior alternative to primary rhesus macaque kidney (pRhMK) cells for detecting various viruses, improving research reliability.
Area of Science:
- Virology
- Cell Biology
- Infectious Disease Research
Background:
- Primary rhesus macaque kidney (pRhMK) cells are crucial for virus detection but present challenges.
- These challenges include potential endogenous viruses, ethical concerns regarding primate use, and inherent variability in primary cultures.
Purpose of the Study:
- To develop a continuous cell line or co-culture system to replace the reliance on pRhMK cells.
- To establish a more reliable and ethical cell culture model for virological studies.
Main Methods:
- Mixed cell monolayers were prepared using Calu-3 and A-549 cell lines.
- These mixed cell cultures were compared against pRhMK cells for their efficacy in detecting a panel of viruses.
- Viruses tested included respiratory viruses, measles, mumps, enteroviruses, and herpes viruses, using both clinical and laboratory strains.
Main Results:
- Calu-3/A-549 cells demonstrated higher sensitivity than pRhMK cells for detecting adenovirus, enteroviruses, and herpes simplex virus.
- The sensitivity of Calu-3/A-549 cells was comparable to pRhMK cells for other respiratory viruses, measles, mumps, and varicella-zoster virus.
Conclusions:
- The Calu-3/A-549 mixed cell culture system serves as an effective alternative to pRhMK cells.
- This novel cell culture approach is suitable for the detection of numerous clinically significant viruses, offering improved research capabilities.
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