Faculty of Medicine, Center of Multidisciplinary Technology for Advanced Medicine (CMUTEAM), Chiang Mai University, 110 Intawaroros Road, Si Phum, Muang, Chiang Mai, 50200, Thailand.
Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand.
Sci Rep. 2024 May 14;14(1):11056. doi: 10.1038/s41598-024-60628-z.
Osteosarcoma is the most common malignant bone cancer in pediatric patients. Patients who respond poorly to chemotherapy experience worse clinical outcomes with a high mortality rate. The major challenge is the lack of effective drugs for these patients. To introduce new drugs for clinical approval, preclinical studies based on in vitro models must demonstrate the potency of the tested drugs, enabling the drugs to enter phase 1 clinical trials. Patient-derived cell culture is a promising testing platform for in vitro studies, as they more accurately recapitulate cancer states and genetic profiles compared to cell lines. In the present study, we established patient-derived osteosarcoma cells (PDC) from a patient who had previously been diagnosed with retinoblastoma. We identified a new variant of a germline mutation in the RB1 gene in the tissue of the patient. The biological effects of this PDC were studied to observe whether the cryopreserved PDC retained a feature of fresh PDC. The cryopreserved PDC preserved the key biological effects, including cell growth, invasive capability, migration, and mineralization, that define the conserved phenotypes compared to fresh PDC. From whole genome sequencing analysis of osteosarcoma tissue and patient-derived cells, we found that cryopreserved PDC was a minor population in the origin tissue and was selectively grown under the culture conditions. The cryopreserved PDC has a high resistance to conventional chemotherapy. This study demonstrated that the established cryopreserved PDC has the aggressive characteristics of osteosarcoma, in particular the chemoresistance phenotype that might be used for further investigation in the chemoresistant mechanism of osteosarcoma. In conclusion, the approach we applied for primary cell culture might be a promising method to generate in vitro models for functional testing of osteosarcoma.
骨肉瘤是儿童中最常见的恶性骨癌。对化疗反应不佳的患者临床预后较差，死亡率较高。主要的挑战是缺乏这些患者的有效药物。为了将新药引入临床批准，基于体外模型的临床前研究必须证明所测试药物的效力，使药物能够进入 1 期临床试验。患者来源的细胞培养是体外研究的有前途的测试平台，因为与细胞系相比，它们更准确地再现了癌症状态和遗传特征。在本研究中，我们从一名先前被诊断患有视网膜母细胞瘤的患者中建立了骨肉瘤患者来源的细胞（PDC）。我们在患者的组织中鉴定了 RB1 基因中种系突变的一个新变体。研究了这种 PDC 的生物学效应，以观察冷冻保存的 PDC 是否保留了新鲜 PDC 的特征。与新鲜 PDC 相比，冷冻保存的 PDC 保留了定义保守表型的关键生物学效应，包括细胞生长、侵袭能力、迁移和矿化。通过对骨肉瘤组织和患者来源细胞的全基因组测序分析，我们发现冷冻保存的 PDC 在起源组织中是一个小群体，并且在培养条件下被选择性生长。冷冻保存的 PDC 对常规化疗具有高抗性。这项研究表明，所建立的冷冻保存的 PDC 具有骨肉瘤的侵袭特征，特别是耐药表型，可用于进一步研究骨肉瘤的耐药机制。总之，我们应用于原代细胞培养的方法可能是一种有前途的方法，可用于骨肉瘤功能测试的体外模型生成。
