Barton Jack, Pacey Katherine, Jain Neha, Kasia Tessa, Edwards Darren, Thevanesan Christine, Straathof Karin, Barone Giuseppe, Anderson John
Cancer Section, DBC Programme, UCL Great Ormond Street Institute of Child Health, London, UK.
Oncology, Great Ormond Street Hospital for Children, London, UK.
F1000Res. 2019 Jun 10;8:823. doi: 10.12688/f1000research.18209.1. eCollection 2019.
: Primary cell culture using serum free media supplemented with growth factors has been used in a number of cancers to propagate primary cells with stem like properties, which form as spherical cellular aggregates. : We systematically evaluated the capacity of freshly disaggregated neuroblastoma tumors to become established as neurospheres in stem cell media using a uniform protocol. 67 primary neuroblastoma samples from patients treated at a single institution were prospectively evaluated for their ability to become established in culture. Samples, either solid tissue or cells from surgical transit fluid both post chemotherapy and chemotherapy naïve, were evaluated from diagnostic needle biopsies or surgical resections. : Overall 37 neurosphere cultures were successfully established from 67 samples. In 11 out of 14 cases investigated by flow cytometry, uniform staining for neuroblastoma markers CD56 and GD2 was demonstrated in CD45 negative non-hemopoietic cells, confirming neuroblastoma origin. : We present a simple and reproducible approach for producing primary neurospheres from neuroblastoma samples, which provides a reliable resource for future work including genetic analysis, stem cell research and models for therapeutics.
使用添加生长因子的无血清培养基进行原代细胞培养已被用于多种癌症，以培养具有干细胞样特性的原代细胞，这些细胞形成球形细胞聚集体。我们使用统一方案系统评估了新鲜解离的神经母细胞瘤肿瘤在干细胞培养基中形成神经球的能力。对来自单一机构接受治疗的患者的67个原发性神经母细胞瘤样本进行前瞻性评估，以确定它们在培养中形成神经球的能力。样本包括实体组织或化疗后及未接受化疗的手术冲洗液中的细胞，这些样本均取自诊断性针吸活检或手术切除组织。总体而言，从67个样本中成功建立了37个神经球培养物。在通过流式细胞术研究的14例病例中的11例中，在CD45阴性的非造血细胞中显示出神经母细胞瘤标志物CD56和GD2的均匀染色，证实了神经母细胞瘤的起源。我们提出了一种从神经母细胞瘤样本中产生原代神经球的简单且可重复的方法，这为包括遗传分析、干细胞研究和治疗模型在内的未来工作提供了可靠的资源。
