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

被引:13
作者
Martinez, Alicia K. Fleming [1 ]
Storz, Peter [1 ]
机构
[1] Mayo Clin Florida, Dept Canc Biol, Jacksonville, FL 32224 USA
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2019年 / 144期
关键词
Cancer Research; Issue; 144; Primary acinar cells; acinar-to-ductal metaplasia; 3-dimensional culture; pancreatic cancer; adenoviral vectors; lentiviral vectors; MUTANT KRAS; ADENOCARCINOMA; NEOPLASIA; NOTCH;
D O I
10.3791/59096
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页数:6
相关论文
共 16 条
[1]  
Artym Vira V, 2010, Curr Protoc Cell Biol, VChapter 10, DOI 10.1002/0471143030.cb1018s48
[2]   A novel explant outgrowth culture model for mouse pancreatic acinar cells with long-term maintenance of secretory phenotype [J].
Blauer, Merja ;
Nordback, Isto ;
Sand, Juhani ;
Laukkarinen, Johanna .
EUROPEAN JOURNAL OF CELL BIOLOGY, 2011, 90 (12) :1052-1060
[3]  
CALDAS C, 1995, INT J PANCREATOL, V18, P1
[4]   Revealing the Cytoskeletal Organization of Invasive Cancer Cells in 3D [J].
Geraldo, Sara ;
Simon, Anthony ;
Vignjevic, Danijela M. .
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2013, (80) :e50763
[5]   Isolation and Culture of Mouse Primary Pancreatic Acinar Cells [J].
Gout, Johann ;
Pommier, Roxane M. ;
Vincent, David F. ;
Kaniewski, Bastien ;
Martel, Sylvie ;
Valcourt, Ulrich ;
Bartholin, Laurent .
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2013, (78) :e50514
[6]  
Grippo PJ, 2003, CANCER RES, V63, P2016
[7]   Spontaneous induction of murine pancreatic intraepithelial neoplasia (mPanIN) by acinar cell targeting of oncogenic Kras in adult mice [J].
Habbe, Nils ;
Shi, Guanglu ;
Meguid, Robert A. ;
Fendrich, Volker ;
Esni, Farzad ;
Chen, Huiping ;
Feldmann, Georg ;
Stoffers, Doris A. ;
Konieczny, Stephen F. ;
Leach, Steven D. ;
Maitra, Anirban .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (48) :18913-18918
[8]   The costs of using unauthenticated, over-passaged cell lines: how much more data do we need? [J].
Hughes, Peyton ;
Marshall, Damian ;
Reid, Yvonne ;
Parkes, Helen ;
Gelber, Cohava .
BIOTECHNIQUES, 2007, 43 (05) :575-+
[9]   Notch and Kras reprogram pancreatic acinar cells to ductal intraepithelial neoplasia [J].
Jean-Paul, De La O. ;
Emerson, Lyska L. ;
Goodman, Jessica L. ;
Froebe, Scott C. ;
Illum, Benjamin E. ;
Curtis, Andrew B. ;
Murtaugh, L. Charles .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (48) :18907-18912
[10]   Identification of Sox9-Dependent Acinar-to-Ductal Reprogramming as the Principal Mechanism for Initiation of Pancreatic Ductal Adenocarcinoma [J].
Kopp, Janel L. ;
von Figura, Guido ;
Mayes, Erin ;
Liu, Fen-Fen ;
Dubois, Claire L. ;
Morris, John P. ;
Pan, Fong Cheng ;
Akiyama, Haruhiko ;
Wright, Christopher V. E. ;
Jensen, Kristin ;
Hebrok, Matthias ;
Sander, Maike .
CANCER CELL, 2012, 22 (06) :737-750