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Updated: Jun 10, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
[Study of infective efficiency of two adenovirus vectors to rBMSC and its mechanism]
Shuo Hu1, Ling-jing Xiong, Jian-ying Yu
1Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, China.
Objective:
To investigate the infective efficiency of Ad5/EGFP versus AdF35/EGFP to rat bone marrow mesenchymal stem cells (rBMSC) and understand its mechanism.
Methods:
rBMSC were isolated from rat bone marrow by density gradient method. The expressions of CD90, CD29, CD44, CD34, CD11b and CD45 on the surface of rBMSC were measured by flow cytometry. The EGFP-carrying Ad5 and AdF35 were infected into rBMSC respectively. The effect of Ad5/EGFP and AdF35/EGFP with different MOIs to rBMSC was tested by MTT. Infected rBMSC were observed by fluorescence microscopy. The infective efficiency and mean fluorescence intensity were determined by flow cytometry after a cellular infection for 1 - 9 days. The expressions of CAR and CD46 on rBMSC were measured by real-time PCR.
Results:
The expressions of CD90, CD29 and CD44 on rBMSC were positive while those of CD34, CD11b and CD45 were negative. When MOI
Conclusion:
The infective efficiency of Ad5/EGFP is significantly higher than that of AdF35/EGFP. It may explain the expression patterns of CAR and CD46 on rBMSC. AdF35 is not a suitable gene-carrying vector to infect rBMSC. Yet Ad5 can be an efficient vector carrying target gene to infect rBMSC for curing diseases.
Insights
Adenovirus serotype 5 (Ad5)/EGFP shows significantly higher infectivity in rat bone marrow mesenchymal stem cells (rBMSC) compared to AdF35/EGFP. Ad5 is a suitable vector for gene delivery to rBMSC, unlike AdF35.
Area of Science:
- Gene therapy vectors
- Stem cell biology
- Adenovirus research
Context:
- Rat bone marrow mesenchymal stem cells (rBMSC) are crucial for regenerative medicine.
- Adenoviral vectors are widely used for gene delivery.
- Understanding vector efficiency in stem cells is vital for therapeutic applications.
Purpose:
- To compare the infective efficiency of Ad5/EGFP and AdF35/EGFP in rBMSC.
- To elucidate the underlying mechanisms of differential infectivity.
- To determine the suitability of Ad5 and AdF35 as gene-carrying vectors for rBMSC.
Summary:
- rBMSC expressed CD90, CD29, and CD44, with low CD46 and high Coxsackie-adenovirus receptor (CAR) expression.
- Ad5/EGFP demonstrated significantly higher infectivity and expression in rBMSC compared to AdF35/EGFP.
- Infection with Ad5/EGFP did not inhibit rBMSC growth, suggesting Ad5 is a promising vector for rBMSC gene delivery.
Impact:
- Ad5/EGFP is a more efficient vector for gene delivery to rBMSC than AdF35/EGFP.
- The findings provide a mechanistic basis for vector selection in rBMSC-based therapies.
- Ad5 holds potential as an effective gene-carrying vector for treating diseases using rBMSC.
