Department of Anatomical Sciences, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Cellular and Molecular Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Iran J Med Sci. 2024 Sep 1;49(9):590-603. doi: 10.30476/ijms.2023.99642.3175. eCollection 2024 Sep.
Researchers are looking for a way to improve the myogenic differentiation of stem cells. Adipose-derived stem cells (ADSCs), known for their multipotency and regenerative capabilities, have been extensively studied for their therapeutic potential. Meanwhile, PC12 cells, derived from rat pheochromocytoma, have been found pivotal in neuroscience research, particularly as a neuronal model system. The current study investigated the effect of the PC12 adrenal pheochromocytoma cell line on the myogenic differentiation of ADSCs.
This experimental study was conducted during 2019-2022 (Ahvaz, Iran). Differentiation of ADSCs was induced by using 3 μg/mL 5-azacytidine for 24 hours. Then, the culture media was changed with Dulbecco's Modified Eagle-High Glucose (DMEM-HG) containing 5% horse serum (HS) and kept for 7 days. Different percentages of differentiated ADSCs and PC12 (100:0, 70:30, 50:50, 30:70) were cocultured for 7 days in DMEM-HG containing 5% HS. PC12 was labeled with cell tracker C7000. The real-time polymerase chain reaction and Western blotting techniques were utilized to assess gene and protein expression. All experiments were repeated three times. Data were analyzed using GraphPad Prism 8.0.2 software with a one-way analysis of variance. P<0.05 was considered statistically significant.
PC12 visualization confirmed the accuracy of the co-culture process. The differentiated cells showed an aligned, multinucleated shape. The differentiated ADSCs revealed significantly elevated levels of , , and gene expression compared with undifferentiated ADSCs (P<0.0001). The ADSCs cocultured with PC12 cells showed significantly higher , , and gene expression than differentiated ADSCs (P<0.001). ADSCs cocultured with 50% PC12 revealed significantly higher MYH and nAchR protein expression than the differentiated group (P<0.01 and P<0.001).
Coculturing PC12 cells and ADSCs improves the efficiency of myogenic differentiation. However, the effectiveness of myogenic differentiation depends on the proportions of administered PC12 cells.
研究人员正在寻找一种方法来提高干细胞的成肌分化。脂肪来源的干细胞（ADSCs）因其多能性和再生能力而被广泛研究，具有治疗潜力。同时，源自大鼠嗜铬细胞瘤的 PC12 细胞在神经科学研究中被发现是至关重要的，特别是作为神经元模型系统。本研究探讨了 PC12 肾上腺嗜铬细胞瘤细胞系对 ADSC 成肌分化的影响。
这是一项 2019-2022 年在伊朗阿瓦兹进行的实验研究。通过使用 3μg/mL 5-氮杂胞苷诱导 ADSC 分化 24 小时。然后，将含有 5%马血清（HS）的 Dulbecco's Modified Eagle-High Glucose（DMEM-HG）培养基更换，并保持 7 天。不同百分比的分化 ADSC 和 PC12（100:0、70:30、50:50、30:70）在含有 5%HS 的 DMEM-HG 中共同培养 7 天。用细胞追踪剂 C7000 标记 PC12。利用实时聚合酶链反应和 Western 印迹技术评估基因和蛋白表达。所有实验均重复 3 次。使用 GraphPad Prism 8.0.2 软件进行数据分析，采用单向方差分析。P<0.05 被认为具有统计学意义。
PC12 的可视化证实了共培养过程的准确性。分化后的细胞呈现出对齐的多核形状。与未分化的 ADSC 相比，分化的 ADSC 显示出 、 和 基因表达水平显著升高（P<0.0001）。与分化的 ADSC 相比，与 PC12 细胞共培养的 ADSC 显示出 、 和 基因表达水平显著升高（P<0.001）。与分化的 ADSC 相比，与 50%PC12 共培养的 ADSC 显示出更高的 MYH 和 nAchR 蛋白表达（P<0.01 和 P<0.001）。
共培养 PC12 细胞和 ADSC 可提高成肌分化效率。然而，成肌分化的效率取决于给予的 PC12 细胞的比例。
