There are numerous approaches to improve the low transduction efficiency of retroviral vectors in two-dimensional (2D) cell culture substrates. However, the effect of a three-dimensional (3D) microenvironment, which better mimics in vivo conditions, is unknown. Cytocompatible hyaluronic acid (HA) hydrogels are a good candidate to study this issue. Here, photocrosslinkable HA hydrogels with an elastic modulus of 1.0–2.7 kPa were successfully prepared by varying the degree of methacrylation in the HA backbone. Culturing human adipose-derived stem cells (hASCs) in a 3D microenvironment significantly reduces the amount of time required for retroviral gene transduction compared with the conventional 2D method and maintains a high transduction efficiency. This acceleration of retroviral gene transduction correlates with the rate of cell-cycle synchronization. hASCs cultured in a 3D microenvironment have a shorter G1 phase and total cell-cycle length than hASCs cultured using the conventional 2D method. This cell-cycle regulation is dependent on expression of cyclin D1. In summary, prior culturing of hASCs in a 3D microenvironment accelerates retroviral gene transduction by regulating cyclin D1 expression and accelerating cell-cycle synchronization. We conclude that priming via culturing in a 3D microenvironment facilitates efficient and rapid retroviral gene transduction of hASCs without inducing apoptosis.

有许多方法可以提高逆转录病毒载体在二维（2D）细胞培养基质中的低转导效率。但是，更好地模拟体内条件的三维（3D）微环境的效果尚不清楚。细胞相容性透明质酸（HA）水凝胶是研究此问题的理想人选。在这里，通过改变HA主链上的甲基丙烯酸化程度，成功制备了具有1.0-2.7 kPa弹性模量的可光交联的HA水凝胶。与传统的2D方法相比，在3D微环境中培养人类脂肪干细胞（hASC）可以显着减少逆转录病毒基因转导所需的时间，并保持较高的转导效率。逆转录病毒基因转导的这种加速与细胞周期同步的速率有关。与使用常规2D方法培养的hASC相比，在3D微环境中培养的hASC具有更短的G1期和总细胞周期长度。这种细胞周期调节取决于细胞周期蛋白D1的表达。总之，先前在3D微环境中培养hASC可以通过调节细胞周期蛋白D1的表达并加速细胞周期同步来加速逆转录病毒基因的转导。我们得出的结论是，通过在3D微环境中培养进行引物促进了hASC的高效和快速逆转录病毒基因转导，而不会诱导细胞凋亡。这种细胞周期调节取决于细胞周期蛋白D1的表达。总之，在3D微环境中进行hASC的先前培养可通过调节细胞周期蛋白D1的表达并加速细胞周期同步来加速逆转录病毒基因的转导。我们得出的结论是，通过在3D微环境中培养进行引物促进了hASC的高效和快速逆转录病毒基因转导，而不会诱导细胞凋亡。这种细胞周期调节取决于细胞周期蛋白D1的表达。总之，先前在3D微环境中培养hASC可以通过调节细胞周期蛋白D1的表达并加速细胞周期同步来加速逆转录病毒基因的转导。我们得出的结论是，通过在3D微环境中培养进行引物促进了hASC的高效和快速逆转录病毒基因转导，而不会诱导细胞凋亡。