Development of patient derived organoids for cancer drug screening applications

Cancer is a disease characterised by abnormal cell growth that can invade or spread to other regions of the body. Organoids are three-dimensional ex vivo tissue cultures made from embryonic stem cells, induced pluripotent stem cells, progenitor cells or tissue that serve as a physiological model for cancer research. These are designed to recapitulate the in vivo properties of tumours. Importantly, effective recapitulation of the structure of tissues and function is believed to predict patient response, allowing for the creation of personalised therapy in a timely manner that may be used in the clinic. This Review discusses the pre-clinical model and different types of human organoids as models for the development of high throughput drug screening and also aims to highlight how organoids are shaping the future of cancer research.     

人源类器官的研究进展及在口腔医学的展望

类器官是指在体外三维培养构建的,依赖于人造细胞外基质的多细胞团,具有自我更新、自我组织能力,并维持了其来源组织的生理结构和功能。类器官作为一种新兴的研究模型,兼具细胞系和动物模型两者的优点。其构建材料简单、培养效率高、耗时短,在疾病研究模型构建、临床药敏试验和再生医学中有良好的应用前景。类器官模型主要分为三种细胞来源类型：胚胎干细胞/诱导多能干细胞、成体干细胞、肿瘤细胞。本文将重点介绍三种细胞来源类器官最新的研究进展,并展望其在口腔医学中的应用。     

基于生物技术的肠道干细胞研究进展

肠道干细胞对于肠上皮各类细胞稳态的维持十分重要。正常情况下,位于隐窝基底部的肠道干细胞会不断向隐窝顶部迁移,在迁移过程中会分化形成不同的肠上皮细胞。肠道干细胞大约3-5天更新一次以维持肠道上皮的更新和损伤后修复。其中,在肠道上皮完整的条件下维持肠上皮动态平衡的主要是活跃型肠道干细胞,在肠道上皮损伤后发挥修复作用的主要是静止型肠道干细胞。近年来,随着生物技术的发展,关于肠道干细胞的研究一直在不断更新与发展。本文以国内外相关文献为基础,对应用于肠道干细胞研究中的主要生物技术、新发现的肠道干细胞标记物和调控因子进行综述。     

类器官在放射治疗中应用的研究进展

随着核能在军事、科研和医学各个领域的广泛应用,放射医学已成为一门重要的学科。迄今为止,放射医学的研究大多使用细胞模型和动物模型,然而这两种模型都有其局限性,类器官模型的出现填补了这些传统模型留下的空白。类器官是源自组织干细胞的3D体外培养系统,类器官因其具有与原始组织相似的结构、功能和遗传特征而被广泛应用于放射医学、疾病研究、药物开发和癌症建模等领域。本综述按放射治疗部位介绍了各种类器官模型在放射治疗中的研究应用,并对类器官模型目前的局限性及未来前景进行相关论述。     

新型编织型生物反应器内大量乳猪肝细胞的组织化培养

目的:使反应器内培养的肝细胞能在较长时间保持肝细胞的特殊功能和活力,提高生物反应器的功效并为反应器内储存和运输肝细胞提供可能。方法:将新生实验型小猪肝细胞与微载体共同培养,待肝细胞与微载体充分粘附形成微载体-球形聚集体后,将其置入新型编织型生物反应器的外腔,用培养液循环式人工毛细管培养系统进行培养,在培养液内加入氯化氨、醋胺酚检测生物反应器的转化功能,行无血清培养检测肝细胞的白蛋白的合成功能,观察肝细胞的酶漏出量。锥虫蓝染色观察肝细胞的活力,电镜观察其超微结构。结果:生物反应器内培养的肝细胞在1周内能保持较高的活力,并保持着较高的氯化氨、醋胺酚的生物转化及白蛋白合成功能,酶的漏出量也少。肝细胞超微结构示内质网、线粒体等细胞器丰富,核内染色体分布均匀,肝细胞间的微绒毛形成胆小管样结构。结论:肝细胞与微载体及肝细胞之间的聚集,形成直径大小不等的肝细胞-微载体球形聚集体,这种由肝细胞重新结合而成的聚集体类似于体内的肝组织结构,在新型编织型生物反应器内的中空纤维网架的支持下,肝细胞聚集体均匀地分散其中,为肝细胞的生长提供了一个类似体内内环境的三维空间,因而在无氧合器供氧的情况下,肝细胞也能在1周内保持较好的形态和功能。     

Organoids and organ chips in ophthalmology

Recent advances have driven the development of stem cell-derived, self-organizing, three-dimensional miniature organs, termed organoids, which mimic different eye tissues including the retina, cornea, and lens. Organoids and engineered microfluidic organ-on-chips (organ chips) are transformative technologies that show promise in simulating the architectural and functional complexity of native organs. Accordingly, they enable exploration of facets of human disease and development not accurately recapitulated by animal models. Together, these technologies will increase our understanding of the basic physiology of different eye structures, enable us to interrogate unknown aspects of ophthalmic disease pathogenesis, and serve as clinically-relevant surrogates for the evaluation of ocular therapeutics. Both the burden and prevalence of monogenic and multifactorial ophthalmic diseases, which can cause visual impairment or blindness, in the human population warrants a paradigm shift towards organoids and organ chips that can provide sensitive, quantitative, and scalable phenotypic assays. In this article, we review the current situation of organoids and organ chips in ophthalmology and discuss how they can be leveraged for translational applications.     

卵巢癌类器官模型的研究进展和治疗新策略

卵巢癌是全球女性生殖系统三大恶性肿瘤之一,严重威胁着女性的生命健康。近年来,研究者不断改进卵巢癌的治疗手段,但是卵巢癌的5年生存率依然并不乐观。随着精准医疗的推行,类器官模型的临床前研究逐渐进入科研工作者的视野。本文旨在介绍卵巢癌相关实验类器官具有高度保持的肿瘤异质性、模拟肿瘤缺氧微环境、模拟肿瘤的发生转移、药物筛选和精准医学、基因编辑系统和模拟疾病。类器官模型库的建立可以改善卵巢癌治疗发展方向。在3D生物打印和类器官的联合下,有望成为未来药物筛选的新途径。     

5-Aminolevulinic acid-mediated photodynamic activity in patient-derived cholangiocarcinoma organoids

BackgroundAccurate diagnosis of the disease extension of cholangiocarcinoma (CCA) is often difficult in clinical practice. The diagnostic yield of conventional pre-operative imaging or endoscopic procedures is sometimes insufficient for the evaluation of longitudinal spreading of CCA. Here we investigated the usefulness of 5-aminolevulinic acid (5-ALA) for the pre- or intra-operative diagnosis of CCA, using patient-derived organoids.MethodsFour CCA- and two adjacent tissue-derived organoids were established. After 5-ALA treatment, we assessed their photodynamic activity using fluorescence microscopy.ResultsCCA organoids established from different patients showed diverse morphology in contrast to monolayer structures of non-tumor organoids, and had the ability to form subcutaneous tumors in immunodeficient mice. CCA organoids demonstrated remarkably high photodynamic activity based on higher accumulation of protoporphyrin IX as a metabolite of 5-ALA compared to non-tumor organoids (40–71% vs. < 4%, respectively). Importantly, cancer cell-specific high photodynamic activity distinguished the organoids originated from biliary stenotic lesions from those of non-stenotic lesions in a CCA patient. The high photodynamic activity did not depend on the expression profile of heme biosynthesis genes.ConclusionsDistinct 5-ALA-based photodynamic activity could have diagnostic potential for the discrimination of CCA from non-tumor tissues.     

Establishment and characterization of meningioma patient-derived organoid

Meningioma is a central nervous system tumor originated from arachnoid cells. 2D cell culture is widely used as a platform for tumor research as it enables us to culture cells in in vitro and a controlled environment. However, in 2D culture condition, 3D architecture of in vivo tumor mass is lost and phenotypic change may occur. Due to the drawbacks of 2D cell culture, organoid culture is seen as an alternative platform for disease modeling, drug testing and personalized medicine. The objective of this study was to establishing protocol for culturing cells from patient meningioma tissue in in vitro 3D environment. Eight meningiomas were collected for the 3D organoid culture. Cells of 5 meningioma tissues survived and proliferated. Under 3D culture condition, cell aggregates were formed and cytoplasmic processes linking the cell aggregates could be observed. In H&E staining, ovaloid cells and spindle cells were observed. Resembling cultured organoids observed under the light microscope, cell aggregates were also observed in the H&E staining. Epithelial Membrane Antigen (EMA) staining was positive. In 4 (80%) cultured organoids, low Ki67 index (≤6%) were measured. In one cultured organoid, a high Ki67 index (12.8%) was seen. The result of this study revealed the feasibility of culturing meningioma cells in in vitro 3D culture condition. Organoid technology showed its potential as an alternative platform for meningioma research.