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Assessment of Transfection of AdCMV-EGFP to Rat Submandibular Gland Cells

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Abstract

We evaluated the efficiency of transfecting adenoviral vectors encoding enhanced green fluorescent protein (AdCMV-EGFP) into rat submandibular gland cells and the effects of gene transfer on cell proliferation and secretory function. Isolated submandibular gland cells were transfected with different titers (or multiplicity of infection, MOI) of AdCMV-EGFP. The transfection efficiency was evaluated by quantifying EGFP-positive cells by inverted fluorescence microscopy, cell proliferation by MTT assay, and cell secretory activity by measuring α-amylase in culture medium. A transfection efficiency of up to 70.8 % was achieved in submandibular gland cells. MTT assay showed that increased viral titers resulted in significant inhibition of cell proliferation, which occurs on day 5 post-transfection. Simultaneously, the amylase levels started to reduce with a significant decrease on day 7 after transfection. The results show that AdCMV-EGFP transfection of submandibular gland cells at higher MOI results in cytotoxicity, decreased cell proliferation, and secretory function. However, the lower adenoviral titers (e.g., 200 particles/cell) could be an efficient and safe labeling tool for gene transfer to submandibular gland cells.

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Acknowledgments

This work was supported by Grants from the National Natural Science Foundation (No:81271155, 30830108, 81300852), Jiangsu Province Natural Science Foundation of China (BK20130079), Key Project of Science and Technology Bureau of Jiangsu Province(No.BL2013002), the Youth Start Fund of Nan**g City, (No. 2011-19-198*), and The Third Level Fund for the Young Talents in the Health Field of Nan**g City.

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Authors and Affiliations

  1. Department of Cariology and Endodontics, Institue and Hospital of Stomatology, Nan**g University Medical School, Nan**g, 210008, China

    Chao Liu, Leiying Miao, Weibin Sun, **aonan Wu & Fuhua Yan

  2. School of Stomatology, Jilin University, Changchun, 130021, China

    Hongchen Sun

  3. Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, NIH, DHHS, Bethesda, MD, USA

    Changyu Zheng

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  1. Chao Liu
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  2. Leiying Miao
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  3. Weibin Sun
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  4. **aonan Wu
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  5. Fuhua Yan
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  6. Hongchen Sun
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  7. Changyu Zheng
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Correspondence to Leiying Miao or Hongchen Sun.

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Chao Liu and Leiying Miao have contributed equally to this work.

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Liu, C., Miao, L., Sun, W. et al. Assessment of Transfection of AdCMV-EGFP to Rat Submandibular Gland Cells. Cell Biochem Biophys 71, 147–153 (2015). https://doi.org/10.1007/s12013-014-0177-0

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  • DOI: https://doi.org/10.1007/s12013-014-0177-0

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