Enhancement of transduction efficiency using Adeno-associated viral vectors by chemical pretreatment to mice bladder urothelium

Hamada, Akihiro; Kita, Yuki; Murakami, Kimiya; Matsumoto, Keiyu; Sakatani, Toru; Sano, Tsuyoshi; Ogawa, Osamu; Kobayashi, Takashi

doi:10.1016/j.jviromet.2020.113854

Cited by 5 publications

(2 citation statements)

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“…Hamada et al Published a methodological study to optimize the transfection e ciency of AAV in mouse bladder. They thought that mice treated with 0.1 N HCl for 4 min had the highest survival rates and had nearly 14.5% transfection e ciency [23]. We conducted an updated study and found that although the rat bladder treated with HCl could signi cantly improve the transfection e ciency compared with the control group, the transfection e ciency of AAV treated with 0.1N NaOH 4 min could get the best transfection effect.…”

Section: Discussionmentioning

confidence: 88%

“…We examined the optimal chemical compounds for pretreatment prior to AAV infection. Chemical compounds all brought in sigma used and treatment times were as follows: 0.25% trypsin for 15 min [15], 10 mg/mL protamine sulfate (PS) for 20 min [16], 0.11% poly-L-lysine (PLL) for 15 min [15], 40 IU/mL Hyaluronidase for 20 min [17], 0.1% sodium dodecyl sulfate (SDS) for 15 min [13], 0.1% n-dodecyl-β-dmaltoside (DDM) for 15 min [13], 0.1 N hydrochloric acid (HCl) for 4 min [23], 0.1 N sodium hydroxide (NaOH) for 5 min [18] or PBS for 15 min as a control. All pretreatments were in a total volume of 200 μL.…”

Section: Chemical Pretreatmentmentioning

confidence: 99%

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Optimized Pretreatment Method for Long-Term Transduction Efficiency of AAV Into Rat Bladder

Zhou

Chen

et al. 2022

Preprint

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Objective We sought the optimal preprocessing method to improve the long-term transfer efficiency of adeno-associated viral (AAV) vectors of rat bladder urothelium. Methods Rat bladders were pretreated by eight different chemical reagents including PBS, trypsin, poly-L-lysine (PLL), protamine sulfate (PS), sodium dodecyl sulfate (SDS), n-dodecyl-β-d-maltoside (DDM), HCl or NaOH followed by transurethral instillation with AAV2/9-CMV-zsgreen. The transfection efficiency was evaluated by the percentage of urothelium with green fluorescence after 4 weeks. Results We employed eight different chemical components and eleven pretreatment methods. The rat bladder was harvested and evaluated without any mortality at 4 weeks post-infection. Compared with other treatment groups, the rate of urothelium transfection is comparatively high in the groups of rats treated with HCl or NaOH. In addition, rats treated with 0.1N NaOH 4 min showed significantly highest transfection efficiency (about 3%). The histopathologic results showed regular morphology of the bladder treated with HCl or NAOH and no significant damage was observed. Conclusion Pretreatment with 0.1N NaOH 4 min could improve the transfection efficiency of AAV in bladder urothelium and maintain the transfection efficiency for a long time without pathological damage. Importance We found that pretreatment of the bladder with HCl and NaOH can improve the transfection efficiency of AAV for a long time compared to control group. In addition, rats treated with 0.1N NaOH 4 min could have the most significant effect among these strategies, which was discovered for the first time. This could be valuable for future bladder gene therapy.

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Section: Discussionmentioning

confidence: 88%