Construction of oral squamous cell carcinoma organoids in vitro 3D-culture for drug screening.

IF 2.9 3区 医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE
Oral diseases Pub Date : 2025-01-01 Epub Date: 2024-06-18 DOI:10.1111/odi.15044
Xin-Yuan Zhang, Yi Sui, Xiao-Feng Shan, Lu-Ming Wang, Lei Zhang, Shang Xie, Zhi-Gang Cai
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引用次数: 0

Abstract

Objective: Patient-derived organoids are potent pre-chemotherapy models. Due to limited research on diverse types of oral squamous cell carcinoma (OSCC) and construction efficiency, our goal was to optimize OSCC organoid models from various sites and assess drug responsiveness.

Methods: We screened and optimized culture media, employing three-dimensional techniques to construct human-derived oral squamous cell carcinoma (OSCC) organoid models in vitro. Morphological validation, immunofluorescence analysis, tissue origin verification, and Short Tandem Repeat (STR) sequencing confirmed the consistency between organoids and source tissues. These organoid models were then subjected to varying concentrations of anticancer drugs, with subsequent assessment of cell viability to calculate IC50 values.

Results: Twenty-nine surgical specimens yielded an 86.2% success rate in culturing 25 organoids in vitro. Morphological consistency confirmed nuclear atypia and positive expression of K5, P40, and E-cadherin, indicating squamous epithelial origin. Cultured complex organoids included α-SMA+ tumour-associated fibroblasts and tumour stem cells expressing CD44 and Ki67. STR sequencing affirmed genomic homogeneity between cultured organoids and source tissues. Drug sensitivity testing revealed diverse responses among organoids, highlighting their value for assessing drug sensitivity.

Conclusions: An efficient OSCC organoid culture system for personalized in vitro drug sensitivity screening was established, laying the foundation for precise treatment development.

构建用于药物筛选的体外三维培养口腔鳞状细胞癌器官组织。
目的:源自患者的类器官是有效的化疗前模型。由于对不同类型口腔鳞状细胞癌(OSCC)和构建效率的研究有限,我们的目标是优化来自不同部位的OSCC类器官模型并评估药物反应性:我们筛选并优化了培养基,采用三维技术在体外构建了人源性口腔鳞状细胞癌(OSCC)类器官模型。形态学验证、免疫荧光分析、组织来源验证和短串联重复(STR)测序证实了类器官模型与来源组织之间的一致性。然后,将这些类器官模型置于不同浓度的抗癌药物中,评估细胞存活率,计算 IC50 值:29例手术标本体外培养25个类癌组织的成功率为86.2%。形态学一致性证实了核不典型性以及 K5、P40 和 E-cadherin 的阳性表达,表明其来源于鳞状上皮。培养的复合有机体包括α-SMA+肿瘤相关成纤维细胞和表达CD44和Ki67的肿瘤干细胞。STR 测序证实了培养的有机体与来源组织之间的基因组同质性。药物敏感性测试显示有机体之间存在不同的反应,突出了它们在评估药物敏感性方面的价值:结论:建立了一种高效的OSCC类器官培养系统,可用于个性化体外药物敏感性筛选,为精准治疗的开发奠定了基础。
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来源期刊
Oral diseases
Oral diseases 医学-牙科与口腔外科
CiteScore
7.60
自引率
5.30%
发文量
325
审稿时长
4-8 weeks
期刊介绍: Oral Diseases is a multidisciplinary and international journal with a focus on head and neck disorders, edited by leaders in the field, Professor Giovanni Lodi (Editor-in-Chief, Milan, Italy), Professor Stefano Petti (Deputy Editor, Rome, Italy) and Associate Professor Gulshan Sunavala-Dossabhoy (Deputy Editor, Shreveport, LA, USA). The journal is pre-eminent in oral medicine. Oral Diseases specifically strives to link often-isolated areas of dentistry and medicine through broad-based scholarship that includes well-designed and controlled clinical research, analytical epidemiology, and the translation of basic science in pre-clinical studies. The journal typically publishes articles relevant to many related medical specialties including especially dermatology, gastroenterology, hematology, immunology, infectious diseases, neuropsychiatry, oncology and otolaryngology. The essential requirement is that all submitted research is hypothesis-driven, with significant positive and negative results both welcomed. Equal publication emphasis is placed on etiology, pathogenesis, diagnosis, prevention and treatment.
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