Lv Chenlong, Li Shangkun, Sang Min, Cui Tingting, Xie Jinghui
State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, School of Basic Medical Sciences, Xinjiang Medical University, Urumqi 830000, China.
Xinjiang Key Laboratory of Molecular Biology for Endemic Diseases, School of Basic Medical Sciences, Xinjiang Medical University, Urumqi 830000, China.
Int J Mol Sci. 2025 Jul 4;26(13):6435. doi: 10.3390/ijms26136435.
Stem cells cultured in cell aggregates exhibit higher cell survival rates and enhanced anti-inflammatory and angiogenic effects compared to single cells, constructing a stable and economical cell aggregate culture system that can accurately adjust the mass transfer distance of nutrients, which contributes to improving the therapeutic effects of stem cell aggregates. In this study, an alginate hydrogel microsphere culture system (Alg-HM) was prepared using electrostatic spraying technology and refined by optimizing the electrostatic spraying technology parameters, such as the sodium alginate concentration, voltage, electrospray injection speed, and nozzle inner diameter. Furthermore, by setting the Tip-dropped culture system (Tip-D culture system, created by dropping the resuspended hMSC aggregate-hydrogel solution with a tip to form the hydrogel microsphere) and Matrigel culture system (created by dropping the resuspended hMSC aggregates-Matrigel solution with a tip to form the Matrigel culture system) as the control group and Alg-HM as the experimental group, the culture effect of hMSC aggregates in the optimized Alg-HM culture system was tested; CCK-8 detection and Ki-67 immunofluorescence staining showed that the Alg-HM culture system significantly enhanced the cell proliferation activity of hMSC aggregates after 7 and 14 days of culture. The Calcein-AM/PI cell staining results showed that the Alg-HM culture system can significantly reduce the central necrosis of hMSC aggregates. The RNA sequencing results showed that the Alg-HM culture system can significantly activate the signaling pathways related to cell proliferation in hMSCs. This culture system is helpful for the culture of cell aggregates in vitro and efficient transplantation in vivo.
与单细胞相比，在细胞聚集体中培养的干细胞表现出更高的细胞存活率以及更强的抗炎和促血管生成作用。构建一种稳定且经济的细胞聚集体培养系统，该系统能够精确调节营养物质的传质距离，这有助于提高干细胞聚集体的治疗效果。在本研究中，利用静电喷涂技术制备了藻酸盐水凝胶微球培养系统（Alg-HM），并通过优化静电喷涂技术参数（如海藻酸钠浓度、电压、电喷雾注射速度和喷嘴内径）对其进行了改进。此外，将滴尖培养系统（通过用滴管滴加重悬的人骨髓间充质干细胞聚集体-水凝胶溶液以形成水凝胶微球而创建的滴尖培养系统，Tip-D培养系统）和基质胶培养系统（通过用滴管滴加重悬的人骨髓间充质干细胞聚集体-基质胶溶液以形成基质胶培养系统）设为对照组，将Alg-HM设为实验组，测试了人骨髓间充质干细胞聚集体在优化后的Alg-HM培养系统中的培养效果；CCK-8检测和Ki-67免疫荧光染色显示，Alg-HM培养系统在培养7天和14天后显著增强了人骨髓间充质干细胞聚集体的细胞增殖活性。钙黄绿素-AM/PI细胞染色结果表明，Alg-HM培养系统可显著减少人骨髓间充质干细胞聚集体的中心坏死。RNA测序结果表明，Alg-HM培养系统可显著激活人骨髓间充质干细胞中与细胞增殖相关的信号通路。该培养系统有助于细胞聚集体的体外培养和体内高效移植。
