Mimicking and Manipulating Pancreatic Acinar-to-Ductal Metaplasia in 3-dimensional Cell Culture

Alicia K. Fleming Martinez, Peter Storz

Research output: Contribution to journalArticle

Abstract

The differentiation of acinar cells to ductal cells during pancreatitis and in the early development of pancreatic cancer is a key process that requires further study. To understand the mechanisms regulating acinar-to-ductal metaplasia (ADM), ex vivo 3D culture and differentiation of primary acinar cells to ductal cells offers many advantages over other systems. With the technique herein, modulation of protein expression is simple and quick, requiring only one day to isolate, stimulate or virally infect, and begin culturing primary acinar cells to investigate the ADM process. In contrast to using basement membrane matrix, the seeding of acinar cell clusters in collagen I extracellular matrix, allows acinar cells to retain their acinar identity before manipulation. This is vital when testing the contribution of various components to the induction of ADM. Not only are the effects of cytokines or other ectopically administered factors testable through this technique, but the contribution of common mutations, increased protein expression, or knockdown of protein expression is testable via viral infection of primary acinar cells, using adenoviral or lentiviral vectors. Moreover, cells can be re-isolated from collagen or basement membrane matrix at the endpoint and analyzed for protein expression.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number144
DOIs
StatePublished - Feb 11 2019

Fingerprint

Acinar Cells
Metaplasia
Cell culture
Cell Culture Techniques
Proteins
Collagen
Basement Membrane
Modulation
Virus Diseases
Cytokines
Pancreatic Neoplasms
Pancreatitis
Extracellular Matrix
Testing
Mutation

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Mimicking and Manipulating Pancreatic Acinar-to-Ductal Metaplasia in 3-dimensional Cell Culture. / Fleming Martinez, Alicia K.; Storz, Peter.

In: Journal of visualized experiments : JoVE, No. 144, 11.02.2019.

Research output: Contribution to journalArticle

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