从肿瘤制备类器官:3D 体外肿瘤模型

肿瘤组织切成小块。将碎片转移到装有消化缓冲液的试管中。细胞周围的胶原纤维被分解，降解组织的基质。然后，沉淀消化的片段并丢弃大部分消化缓冲液。轻弹管子以打碎颗粒。

该溶液现在由不同大小的细胞团组成。将细胞团块重悬于胰蛋白酶和ROCK抑制剂的溶液中。胰蛋白酶继续分解肿瘤，抑制剂防止细胞死亡。定期移液以打碎团块并分散细胞。然后，将细胞以适当的浓度重悬于基质溶液中。

溶液滴入培养板中以形成基质圆顶。将板倒置在培养箱中，让基质完全凝固。然后，加入细胞培养基并孵育，正面朝上。该基质提供了一个支架来支持类器官的三维生长。

切碎后，将肿瘤块放入15毫升消化缓冲液管中，在37摄氏度下消化1.5至2小时。在消化结束时，通过离心沉淀消化的组织并丢弃上清液。

弹管子以松开细胞沉淀。将细胞重悬于补充有10微摩尔Y-27632 ROCK抑制剂的1毫升预热胰蛋白酶中，在37摄氏度下5分钟。

孵育结束时，用标准 P1000 移液器吸头将组织浆液研磨五次。将试管放回水浴中，再孵育和研磨 5 分钟。

对于基质圆顶铺板，在 9 毫升冷培养基中洗涤消化的细胞，然后通过离心收集细胞。将细胞重悬于1毫升新鲜培养基中进行计数，计数后，再次通过离心收集细胞。在适当体积的基质中稀释肿瘤细胞。

小心地将 200 微升基质细胞溶液滴入 55 摄氏度加热的 6 孔板的每个孔中。让圆顶在室温下凝固 2 分钟，然后将板倒置在 37 摄氏度的培养箱中 20 分钟。

当圆顶完全凝固时，向每个孔中加入2毫升补充有雄激素R1881和ROCK抑制剂的前列腺类器官培养基。将板放回细胞培养箱中。

• In vitro tumor models - Methods for studying tumor behavior in lab-controlled conditions.
• PeproTech Noggin - A protein used to regulate cellular processes in tissue culture.
• Tumor model - A method or process used to replicate tumor environment and behavior for scientific studies.
• Cell pellet - Cluster of cells collected at the bottom of a tube after centrifugation.
• ROCK inhibitor organoids - Compounds that prevent cell apoptosis during organoid creation.
• Noggin conditioned media - Media enriched with Noggin protein, used in cellular culture and experiments.
• Cell pellet in tube - Term refers to cells that have been concentrated by centrifugation in a sample tube.

• Introduce In vitro tumor models by defining this as a replicate of tumor behavior (e.g., in vitro tumor models).
• Key Concepts by summarizing principles involved in cell pellet creation (e.g., cell pellet).
• Underlying Mechanisms through visualizing the process of using ROCK inhibitor in organoid creation.
• Connect the process of tumor tissue mincing to the creation of a cell pellet and suspension in a matrix solution.

• What are in vitro tumor models and how to create a cell pellet in lab conditions?
• What role does the PeproTech Noggin play in the tumor model creation?
• How does the ROCK inhibitor assist in the creation of organoids?

• Practical Applications - Tumor models help study cancer growth and potential cures (e.g., in vitro tumor models).
• Industry Impact - Lab studies benefit the healthcare industry and the field of oncology (e.g., ROCK inhibitor organoids).
• Societal Importance - The research contributes to advancements in cancer treatment options (e.g., Noggin conditioned media).
• Link to Scientific Advancements - Understanding tumor behavior facilitates innovative treatment approaches.