Institute of Cellular Biology and Pathology "N. Simionescu", 8, B.P. Hasdeu Street, 050568 Bucharest, Romania.
Int J Mol Sci. 2021 Jan 9;22(2):598. doi: 10.3390/ijms22020598.
Adenoviral vectors are important vehicles for delivering therapeutic genes into mammalian cells. However, the yield of the adenoviral transduction of murine mesenchymal stromal cells (MSC) is low. Here, we aimed to improve the adenoviral transduction efficiency of bone marrow-derived MSC. Our data showed that among all the potential transduction boosters that we tested, the K2 Transfection System (K2TS) greatly increased the transduction efficiency. After optimization of both K2TS components, the yield of the adenoviral transduction increased from 18% to 96% for non-obese diabetic (NOD)-derived MSC, from 30% to 86% for C57BL/6-derived MSC, and from 0.6% to 63% for BALB/c-derived MSC, when 250 transduction units/cell were used. We found that MSC derived from these mouse strains expressed different levels of the coxsackievirus and adenovirus receptors (MSC from C57BL/6≥NOD>>>BALB/c). K2TS did not increase the level of the receptor expression, but desensitized the cells to foreign DNA and facilitated the virus entry into the cell. The expression of Stem cells antigen-1 (Sca-1) and 5'-nucleotidase (CD73) MSC markers, the adipogenic and osteogenic differentiation potential, and the immunosuppressive capacity were preserved after the adenoviral transduction of MSC in the presence of the K2TS. In conclusion, K2TS significantly enhanced the adenoviral transduction of MSC, without interfering with their main characteristics and properties.
腺病毒载体是将治疗基因递送入哺乳动物细胞的重要载体。然而,腺病毒对鼠间充质基质细胞(MSC)的转导效率较低。在此,我们旨在提高骨髓来源的 MSC 的腺病毒转导效率。我们的数据表明,在我们测试的所有潜在转导增强剂中,K2 转染系统(K2TS)极大地提高了转导效率。在优化 K2TS 成分后,当使用 250 个转导单位/细胞时,腺病毒转导的产量从 NOD 衍生的 MSC 的 18%增加到 96%,从 C57BL/6 衍生的 MSC 的 30%增加到 86%,从 BALB/c 衍生的 MSC 的 0.6%增加到 63%。我们发现这些小鼠品系来源的 MSC 表达不同水平的柯萨奇病毒和腺病毒受体(C57BL/6 衍生的 MSC≥NOD>>>BALB/c)。K2TS 并未增加受体表达水平,但使细胞对外来 DNA 脱敏,并促进病毒进入细胞。在 K2TS 存在下,MSC 的腺病毒转导后,其干细胞抗原-1(Sca-1)和 5'-核苷酸酶(CD73)MSC 标志物的表达、成脂和成骨分化潜能以及免疫抑制能力得以保留。总之,K2TS 显著增强了 MSC 的腺病毒转导,而不干扰其主要特征和性质。