Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing 400715, China.
Int J Mol Sci. 2023 Jul 26;24(15):11967. doi: 10.3390/ijms241511967.
Cells with various structures and proteins naturally come together to cooperate in vivo. This study used cell spheroids cultured in agarose micro-wells as a 3D model to study the movement of cells or spheroids toward other spheroids. The formation dynamics of tumor spheroids and the interactions of two batches of cells in the agarose micro-wells were studied. The results showed that a concave bottom micro-well (diameter: 2 mm, depth: 2 mm) prepared from 3% agarose could be used to study the interaction of two batches of cells. The initial tumor cell numbers from 5 × 10 cells/well to 6 × 10 cells/well all could form 3D spheroids after 3 days of incubation. Adding the second batch of DU 145 cells to the existing DU 145 spheroid resulted in the formation of satellite cell spheroids around the existing parental tumor spheroid. Complete fusion of two generation cell spheroids was observed when the parental spheroids were formed from 1 × 10 and 2 × 10 cells, and the second batch of cells was 5 × 10 per well. A higher amount of the second batch of cells (1 × 10 cell/well) led to the formation of independent satellite spheroids after 48 h of co-culture, suggesting the behavior of the second batch of cells towards existing parental spheroids depended on various factors, such as the volume of the parental spheroids and the number of the second batch cells. The interactions between the tumor spheroids and Human Umbilical Vein Endothelial Cells (HUVECs) were modeled on concave agarose micro-wells. The HUVECs (3 × 10 cell/well) were observed to gather around the parental tumor spheroids formed from 1 × 10, 2 × 10, and 3 × 10 cells per well rather than aggregate on their own to form HUVEC spheroids. This study highlights the importance of analyzing the biological properties of cells before designing experimental procedures for the sequential fusion of cell spheroids. The study further emphasizes the significant roles that cell density and the volume of the spheroids play in determining the location and movement of cells.
细胞与各种结构和蛋白质自然地结合在一起，在体内进行合作。本研究使用琼脂糖微井中培养的细胞球作为 3D 模型，研究细胞或细胞球向其他细胞球的运动。研究了肿瘤细胞球的形成动力学和琼脂糖微井中两批细胞的相互作用。结果表明，由 3%琼脂糖制备的直径为 2mm、深度为 2mm 的凹底微井可用于研究两批细胞的相互作用。初始肿瘤细胞数从 5×10 个/孔到 6×10 个/孔，在孵育 3 天后都能形成 3D 细胞球。在现有的 DU 145 肿瘤细胞球中添加第二批 DU 145 细胞，导致卫星细胞球围绕现有的亲代肿瘤细胞球形成。当亲代细胞球由 1×10 和 2×10 个细胞形成，第二批细胞每孔为 5×10 个时，观察到两代细胞球的完全融合。当第二批细胞的数量较高（每孔 1×10 个细胞）时，在共培养 48 小时后，会形成独立的卫星细胞球，这表明第二批细胞的行为取决于各种因素，如亲代细胞球的体积和第二批细胞的数量。在凹形琼脂糖微井中模拟了肿瘤细胞球与人脐静脉内皮细胞（HUVEC）的相互作用。观察到 HUVEC（3×10 个细胞/孔）聚集在亲代肿瘤细胞球周围，这些细胞球由每孔 1×10、2×10 和 3×10 个细胞形成，而不是自行聚集形成 HUVEC 球。本研究强调了在设计细胞球序贯融合的实验程序之前，分析细胞生物学特性的重要性。研究进一步强调了细胞密度和细胞球体积在决定细胞位置和运动方面的重要作用。
