Biphenotypic Differentiation of Pancreatic Cancer in 3-Dimensional Culture

Yoshihisa Matsushita, Barbara Smith, Michael Delannoy, Maria A. Trujillo, Peter Chianchiano, Ross McMillan, Hirohiko Kamiyama, Hong Liang, Elizabeth D. Thompson, Ralph H. Hruban, William Matsui, Laura D. Wood, Nicholas J. Roberts, James R. Eshleman

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Objective Pancreatic ductal adenocarcinoma (PDAC) is the third most common cause of cancer death in the United States. Improved characterized models of PDAC are needed for drug screening. Methods We grew 4 established pancreatic cancer cell lines in hanging drop cultures to produce spheroids. We also grew organoids from explanted xenografted PDAC and surgically resected primary PDAC. We performed transmission and scanning electron microscopy and compared findings with those of the normal pancreatic duct. We also performed single-cell cloning to determine the potential options for differentiation. Results Spheroids contained tight junctions and desmosomes but lacked zymogen granules, as expected. The former features were present in normal pancreatic duct but absent from PDAC cell lines grown in standard 2-dimensional culture. Spheroids functionally excluded macromolecules in whole mounts. Cells on the surface of PDAC spheroids were carpeted by microvilli except for rare cells with prominent stereocilia. Carpets of microvilli were also seen in low passage organoids produced from xenografts and surgically resected human PDAC, in addition to normal human pancreatic duct. We performed single-cell cloning and resulting spheroids produced both cell phenotypes at the same approximate ratios as those from bulk cultures. Conclusions Pancreatic cancer spheroids/organoids are capable of biphenotypic differentiation.

Original languageEnglish (US)
Pages (from-to)1225-1231
Number of pages7
JournalPancreas
Volume48
Issue number9
DOIs
StatePublished - Oct 1 2019

Keywords

  • PDAC
  • biphenotypic differentiation
  • microvilli
  • organoids
  • spheroids
  • stereocilia

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

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