Nath Shubhankar, Moore Kaitlin
Wellman Center for Photomedicine, Massachusetts General Hospital-Harvard Medical School, Boston, USA.
Bouve College of Health Sciences, Northeastern University, Boston, USA.
Bio Protoc. 2019 Aug 5;9(15):e3314. doi: 10.21769/BioProtoc.3314.
Photodynamic therapy (PDT), is a clinically-approved light-based anti-cancer treatment modality in which a photoactivatable photosensitizer is irradiated with an appropriate wavelength of light to generate cytotoxic molecules to kill cancer cells. In this article, we describe an PDT protocol using a 3-dimensional (3D) model of ovarian cancer that was established on beds of Matrigel. PDT was performed using a liposomal formulation of verteporfin photosensitizer (Visudyne). The cancer cells were genetically-labeled with the fluorescent protein mCherry to facilitate the evaluation of the treatment response. This protocol is advantageous because the mCherry fluorescence is an indicator of cell viability, eliminating the need for external dyes, which often exhibit limited penetration and diffusion into 3D organoids. Additionally, Visudyne PDT achieves significant tumor-killing efficacy in a 3D model for ovarian cancer.
光动力疗法(PDT)是一种临床批准的基于光的抗癌治疗方式,其中用适当波长的光照射可光激活的光敏剂以产生细胞毒性分子来杀死癌细胞。在本文中,我们描述了一种使用在基质胶床上建立的卵巢癌三维(3D)模型的光动力疗法方案。使用维替泊芬光敏剂(Visudyne)的脂质体制剂进行光动力疗法。癌细胞用荧光蛋白mCherry进行基因标记,以促进对治疗反应的评估。该方案具有优势,因为mCherry荧光是细胞活力的指标,无需外部染料,而外部染料通常在三维类器官中的穿透和扩散有限。此外,Visudyne光动力疗法在卵巢癌的三维模型中实现了显著的肿瘤杀伤效果。