T&R Biofab Co., Ltd., Seongnam-si13487, Republic of Korea.
Department of Dermatology, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul296-12, Korea.
ACS Appl Bio Mater. 2022 Nov 21;5(11):5302-5309. doi: 10.1021/acsabm.2c00710. Epub 2022 Oct 20.
A previous study from our laboratory demonstrated the effects of three-dimensional (3D)-printed collagen scaffolds on the maintenance of cryopreserved patient-derived melanoma explants (PDMEs). However, it remains unknown whether 3D-printed collagen scaffolds (3D-PCSs) can be harmonized with any external culture conditions to increase the growth of cryopreserved PDMEs. In this study, 3D-PCSs were manufactured with a 3DX bioprinter. The 3D-printed collagen scaffold-on-frame construction was loaded with fragments of cryopreserved PDMEs (approximately 1-2 mm). 3D-PCSs loaded with patient-derived melanoma explants (3D-PCS-PDMEs) were incubated using two types of methods: (1) in transwells in the presence of a low concentration of oxygen (transwell-hypoxia method) and (2) using a traditional adherent attached to the bottom flat surface of a standard culture dish (traditional flat condition). In addition, we used six different types of media (DMEM high glucose, MEM α, DMEM/F12, RPMI1640, fibroblast basal medium (FBM), and SBM (stem cell basal medium)) for 7 days. The results reveal that the culture conditions of MEM α, DMEM/F12, and FBM using the transwell-hypoxia method show greater synergic effects on the outgrowth of the 3D-PCS-PDME compared to the traditional flat condition. In addition, the transwell-hypoxia method shows a higher expression of the MMP14 gene and the multidrug-resistant gene product 1 (MDR1) than in the typical culture method. Taken together, our findings suggest that the transwell-hypoxia method could serve as an improved, 3D alternative to animal-free testing that better mimics the skin's microenvironment using PDMEs.
先前，我们实验室的研究表明，三维（3D）打印的胶原蛋白支架可维持低温保存的患者来源黑色素瘤外植体（PDME）。然而，尚不清楚 3D 打印胶原蛋白支架（3D-PCSs）是否可以与任何外部培养条件相协调，以增加低温保存的 PDME 的生长。在这项研究中，使用 3DX 生物打印机制造了 3D-PCSs。将 3D 打印的胶原蛋白支架框架结构加载有低温保存的 PDME 碎片（约 1-2mm）。使用两种方法孵育加载有患者来源黑色素瘤外植体的 3D-PCSs：（1）在存在低氧浓度的 Transwell 中（Transwell 低氧法）和（2）使用传统的附着于标准培养皿底面的附着方法（传统平面条件）。此外，我们使用了六种不同的培养基（DMEM 高葡萄糖、MEM α、DMEM/F12、RPMI1640、成纤维细胞基础培养基（FBM）和 SBM（干细胞基础培养基））进行了 7 天的培养。结果表明，与传统平面条件相比，Transwell 低氧法中使用 MEM α、DMEM/F12 和 FBM 的培养条件对 3D-PCS-PDME 的生长具有更大的协同作用。此外，Transwell 低氧法比典型的培养方法显示出更高的 MMP14 基因和多药耐药基因产物 1（MDR1）的表达。综上所述，我们的研究结果表明，Transwell 低氧法可以作为一种改进的、无动物的 3D 替代方法，使用 PDME 更好地模拟皮肤的微环境。
