The Adrenal Pheochromocytoma Cell Line PC12 Efficiently Promotes the Regeneration Capability of Adipose Tissue-Derived Mesenchymal Stem Cells in Myogenesis: A Particular Approach to Improving Skeletal Muscle Cell Regeneration.

IF 1.6 Q2 MEDICINE, GENERAL & INTERNAL Iranian Journal of Medical Sciences Pub Date : 2024-09-01 DOI:10.30476/ijms.2023.99642.3175
Zeinab Shafiei Seifabadi, Dian Dayer, Seyyed Saeed Azandeh, Mohammad Rashno, Vahid Bayati
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引用次数: 0

Abstract

Background: 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.

Methods: 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.

Results: 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 Myh1, Myh2, and Chrn-α1 gene expression compared with undifferentiated ADSCs (P<0.0001). The ADSCs cocultured with PC12 cells showed significantly higher Myh1, Myh2, and Chrn-α1 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).

Conclusion: 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.

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肾上腺嗜铬细胞瘤细胞系 PC12 有效促进脂肪组织间充质干细胞在肌肉生成中的再生能力:改善骨骼肌细胞再生的特殊方法。
背景:研究人员正在寻找改善干细胞成肌分化的方法。脂肪源性干细胞(ADSCs)以其多潜能性和再生能力而闻名,其治疗潜力已被广泛研究。与此同时,从大鼠嗜铬细胞瘤中提取的 PC12 细胞在神经科学研究中,特别是作为神经元模型系统中,也具有举足轻重的作用。本研究探讨了PC12肾上腺嗜铬细胞瘤细胞系对ADSCs成肌分化的影响:本实验研究于 2019-2022 年期间进行(伊朗阿瓦士)。使用 3 μg/mL 5-azacytidine 诱导 ADSCs 分化 24 小时。然后,用含 5%马血清(HS)的 Dulbecco's Modified Eagle-High Glucose(DMEM-HG)更换培养基并保存 7 天。不同比例的分化 ADSCs 和 PC12(100:0、70:30、50:50、30:70)在含 5% HS 的 DMEM-HG 中共培养 7 天。PC12 用细胞追踪器 C7000 标记。利用实时聚合酶链反应和 Western 印迹技术评估基因和蛋白质的表达。所有实验重复三次。使用 GraphPad Prism 8.0.2 软件对数据进行单因素方差分析。结果PC12 的可视化证实了共培养过程的准确性。分化的细胞呈现出排列整齐的多核形状。与未分化 ADSCs 相比,分化 ADSCs 的 Myh1、Myh2 和 Chrn-α1 基因表达水平明显升高(PMyh1、Myh2 和 Chrn-α1 基因表达水平高于分化 ADSCs):PC12细胞与ADSCs共培养可提高成肌分化的效率。然而,成肌分化的效果取决于所加入的 PC12 细胞的比例。
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来源期刊
Iranian Journal of Medical Sciences
Iranian Journal of Medical Sciences MEDICINE, GENERAL & INTERNAL-
CiteScore
3.20
自引率
0.00%
发文量
84
审稿时长
12 weeks
期刊介绍: The Iranian Journal of Medical Sciences (IJMS) is an international quarterly biomedical publication, which is sponsored by Shiraz University of Medical Sciences. The IJMS intends to provide a scientific medium of com­muni­cation for researchers throughout the globe. The journal welcomes original clinical articles as well as clinically oriented basic science re­search experiences on prevalent diseases in the region and analysis of various regional problems.
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