Creation of Formalin-Fixed, Paraffin-Embedded 3D Lung Cancer Cellular Spheroids for the Optimization of Immunohistochemistry Staining Procedures.
Creation of Formalin-Fixed, Paraffin-Embedded 3D Lung Cancer Cellular Spheroids for the Optimization of Immunohistochemistry Staining Procedures.
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DOI：
10.1007/978-1-0716-1278-1_6
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Santiago-Cardona PG
中科院分区：
文献类型：
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作者：
Cabán-Rivera J;Chardón-Colón C;Pedraza-Torres A;Rodríguez YE;Quiñones-Alvarado R;Santiago-Cardona PG
在精准医疗的时代，重要的治疗决策是由临床信息肿瘤蛋白生物标志物的表达决定的。这些生物标志物可以通过免疫组织化学（IHC）在活检或切除获得的肿瘤组织切片中检测到。与所有实验程序一样，免疫组化的几个步骤需要优化。然而，研究者必须避免使用可能难以获得的有价值的人体活检样本来优化免疫组化程序。理想情况下，有价值的活检样本只应在免疫组化方案优化后进行免疫组化。在本章中，我们描述了利用培养细胞生成的三维细胞球体来优化IHC的过程。在这种方法中，培养的细胞被颗粒化成三维球体，然后像组织样本一样进行处理，即固定、包埋、切片、安装在载玻片上，并像人体组织样本一样用免疫组化染色。这些三维细胞球体具有类似于肿瘤切片的组织样结构和细胞结构，并且细胞和抗原结构都被保留下来。因此，在进行人肿瘤样品的免疫组化染色之前，该方法可用于免疫组化优化。
In an era of precision medicine important treatment decisions are dictated by expression of clinically informative tumor protein biomarkers. These biomarkers can be detected by immunohistochemistry (IHC) performed in tumor tissue sections obtained from biopsies or resections. Like all experimental procedures, IHC needs optimization for several of its steps. However, the investigator must avoid optimizing the IHC procedure using valuable human biopsy samples which may be difficult to obtain. Ideally, valuable biopsy samples should only be subjected to IHC once the IHC protocol has been optimized. In this chapter, we describe a procedure for IHC optimization using tri-dimensional (3D) cellular spheroids created from cultured cells. In this approach, cultured cells are pelleted into 3D spheroids, which are then processed just like a tissue sample, namely, fixed, embedded, sectioned, mounted on slides, and stained with IHC just like a human tissue sample. These 3D cellular spheroids have a tissue-like architecture and cellularity resembling a tumor section, and both cellular and antigen structure are preserved. This method is therefore acceptable for IHC optimization before proceeding to the IHC staining of human tumor samples.