Cancer Res Treat.  2023 Jan;55(1):219-230. 10.4143/crt.2021.1166.

Establishing Patient-Derived Cancer Cell Cultures and Xenografts in Biliary Tract Cancer

• Kang J
• Lee JY
• Lee S
• Kim D
• Lim J
• Jun HR
• Jeon S
• Kim YA
• Park HS
• Kim Kp
• Chun SM
• Lee HJ
• Yoo C

• KMID: 2538006
• DOI: http://doi.org/10.4143/crt.2021.1166

Abstract

Purpose
Biliary tract cancers (BTCs) are rare and show a dismal prognosis with limited treatment options. To improve our understanding of these heterogeneous tumors and develop effective therapeutic agents, suitable preclinical models reflecting diverse tumor characteristics are needed. We established and characterized new patient-derived cancer cell cultures and patient-derived xenograft (PDX) models using malignant ascites from five patients with BTC.
Materials and Methods
Five patient-derived cancer cell cultures and three PDX models derived from malignant ascites of five patients with BTC, AMCBTC-01, -02, -03, -04, and -05, were established. To characterize the models histogenetically and confirm whether characteristics of the primary tumor were maintained, targeted sequencing and histopathological comparison between primary tissue and xenograft tumors were performed.
Results
From malignant ascites of five BTC patients, five patient-derived cancer cell cultures (100% success rate), and three PDXs (60% success rate) were established. The morphological characteristics of three primary xenograft tumors were compared with those of matched primary tumors, and they displayed a similar morphology. The mutated genes in samples (models, primary tumor tissue, or both) from more than one patient were TP53 (n=2), KRAS (n=2), and STK11 (n=2). Overall, the pattern of commonly mutated genes in BTC cell cultures was different from that in commercially available BTC cell lines.
Conclusion
We successfully established the patient-derived cancer cell cultures and xenograft models derived from malignant ascites in BTC patients. These models accompanied by different genetic characteristics from commercially available models will help better understand BTC biology.

Keyword

Ascites; Biliary tract neoplasms; Cancer cell cultures; Patient-derived xenograft; High-throughput nucleotide sequencing

Figure

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  Fig. 1 Flow diagram of patient-derived cancer cell cultures and patient-derived xenografts (PDXs) using malignant ascites from patients with biliary tract cancer (BTC).

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  Fig. 2 Morphology of patient-derived cancer cell cultures when grown in 2D cell culture. Patient-derived cancer cell cultures were imaged by light microscopy.

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  Fig. 3 Comparison of histopathology between three primary tumors and matched primary xenograft tumor (×200, scale bar=200 μm).

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  Fig. 4 Oncoprint of commonly altered genes of patient-derived cancer cell cultures and tissues from primary and patient-derived xenograft tumors.

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  Fig. 5 Single nucleotide polymorphism fingerprint analysis for patient-derived cancer cell cultures and patient-derived xenograft authentication. a)RS1950501 and RS2180770 are located in the intronic region of the RIPK3 and RAD51B genes, respectively. BTC, biliary tract cancer.

Reference

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