Department of General, Visceral and Vascular Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Institute of Pathology, Charité - Universitätsmedizin Berlin, Berlin, Germany.
BMC Cancer. 2020 May 13;20(1):417. doi: 10.1186/s12885-020-06929-8.
Pancreatic cancer remains a fatal disease. Experimental systems are needed for personalized treatment strategies, drug testing and to further understand tumor biology. Cell cultures can serve as an excellent preclinical platform, but their generation remains challenging.
Tumor cells from surgically removed pancreatic ductal adenocarcinoma (PDAC) specimens were cultured under novel protocols. Cellular growth and composition were analyzed and culture conditions were continuously optimized. Characterization of cell cultures and primary tumors was performed via hematoxylin and eosin (HE) and immunofluorescence (IF) staining.
Protocols for two- and three-dimensional PDAC primary cell cultures could successfully be established. Primary cell culture depended on dissociation techniques, growth factor supplementation and extracellular matrix components containing Matrigel being crucial for the transformation to three-dimensional PDAC organoids. The generated cultures showed to be highly resemblant to established PDAC primary cell cultures. HE and IF staining for cell culture and corresponding primary tumor characterization could successfully be performed.
The work presented herein shows novel and effective methods to successfully establish primary PDAC cell cultures in a distinct time frame. Factors contributing to cell growth and differentiation could be identified with important implications for further primary cell culture protocols. The established protocols might serve as novel tools in personalized tumor therapy.
胰腺癌仍然是一种致命的疾病。需要实验系统来制定个性化的治疗策略、药物测试，并进一步了解肿瘤生物学。细胞培养可以作为一个极好的临床前平台，但它们的产生仍然具有挑战性。
从手术切除的胰腺导管腺癌（PDAC）标本中分离肿瘤细胞，根据新方案进行培养。分析细胞的生长和组成，并不断优化培养条件。通过苏木精和伊红（HE）和免疫荧光（IF）染色对细胞培养物和原发肿瘤进行特征分析。
成功建立了二维和三维 PDAC 原代细胞培养方案。原代细胞培养取决于分离技术、生长因子的补充以及含有 Matrigel 的细胞外基质成分，这对于向三维 PDAC 类器官的转化至关重要。所生成的培养物与已建立的 PDAC 原代细胞培养物高度相似。成功地对细胞培养物和相应的原代肿瘤进行了 HE 和 IF 染色。
本文介绍的工作展示了在特定时间框架内成功建立 PDAC 原代细胞培养的新方法。确定了促进细胞生长和分化的因素，这对进一步的原代细胞培养方案具有重要意义。所建立的方案可能成为个体化肿瘤治疗的新工具。
