Application of Concave Microwells to Pancreatic Tumor Spheroids Enabling Anticancer Drug Evaluation in a Clinically Relevant Drug Resistance Model

Sang Eun Yeon, Da Yoon No, Sang Hoon Lee, Suk Woo Nam, Il Hoan Oh, Jaehwi Lee, Hyo Jeong Kuh

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Intrinsic drug resistance of pancreatic ductal adenocarcinoma (PDAC) warrants studies using models that are more clinically relevant for identifying novel resistance mechanisms as well as for drug development. Tumor spheroids (TS) mimic in vivo tumor conditions associated with multicellular resistance and represent a promising model for efficient drug screening, however, pancreatic cancer cells often fail to form spheroids using conventional methods such as liquid overlay. This study describes the induction of TS of human pancreatic cancer cells (Panc-1, Aspc-1, Capan-2) in concave polydimethylsiloxane (PDMS) microwell plates and evaluation of their usefulness as an anticancer efficacy test model. All three cell lines showed TS formation with varying degree of necrosis inside TS. Among these, Panc-1 spheroid with spherical morphology, a rather rough surface, and unique adhesion structures were successfully produced with no notable necrosis in concave microwell plates. Panc-1 TS contained growth factors or enzymes such as TGF-β1, CTGF, and MT1-MMP, and extracellular matrix proteins such as collagen type I, fibronectin, and laminin. Panc-1 cells grown as TS showed changes in stem cell populations and in expression levels of miRNAs that may play roles in chemoresistance. Visualization of drug penetration and detection of viability indicators, such as Ki-67 and MitoSOX, were optimized for TS for quantitative analysis. Water-soluble tetrazolium (MTS) and acid phosphatase (APH) assays were also successfully optimized. Overall, we demonstrated that concave PDMS microwell plates are a novel platform for preparation of TS of weakly aggregating cells and that Panc-1 spheroids may represent a novel three-dimensional model for anti-pancreatic cancer drug screening.

Original languageEnglish
Article numbere73345
JournalPLoS One
Volume8
Issue number9
DOIs
Publication statusPublished - 2013 Sep 10

Fingerprint

drug evaluation
Drug Evaluation
antineoplastic agents
drug resistance
Drug Resistance
Tumors
neoplasms
Pharmaceutical Preparations
pancreatic neoplasms
Neoplasms
Pancreatic Neoplasms
Cells
Preclinical Drug Evaluations
drugs
Necrosis
Screening
necrosis
Matrix Metalloproteinase 14
screening
Extracellular Matrix Proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Application of Concave Microwells to Pancreatic Tumor Spheroids Enabling Anticancer Drug Evaluation in a Clinically Relevant Drug Resistance Model. / Yeon, Sang Eun; No, Da Yoon; Lee, Sang Hoon; Nam, Suk Woo; Oh, Il Hoan; Lee, Jaehwi; Kuh, Hyo Jeong.

In: PLoS One, Vol. 8, No. 9, e73345, 10.09.2013.

Research output: Contribution to journalArticle

Yeon, Sang Eun ; No, Da Yoon ; Lee, Sang Hoon ; Nam, Suk Woo ; Oh, Il Hoan ; Lee, Jaehwi ; Kuh, Hyo Jeong. / Application of Concave Microwells to Pancreatic Tumor Spheroids Enabling Anticancer Drug Evaluation in a Clinically Relevant Drug Resistance Model. In: PLoS One. 2013 ; Vol. 8, No. 9.
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