Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids

Po Hien Ear; Guiying Li; Meng Wu; Ellen Abusada; Andrew M Bellizzi; James R Howe

doi:10.3791/60303

Back

Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids

Journal article

Peer reviewed

Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids

Po Hien Ear, Guiying Li, Meng Wu, Ellen Abusada, Andrew M Bellizzi and James R Howe

Journal of visualized experiments : JoVE, Vol.2019(152), e60303

10/14/2019

DOI: 10.3791/60303

PMCID: PMC7654522

PMID: 31657801

Files and links (1)

url

https://www.ncbi.nlm.nih.gov/pmc/articles/7654522View

Open Access

Abstract

Small bowel neuroendocrine tumors (SBNETs) are rare cancers originating from enterochromaffin cells of the gut. Research in this field has been limited because very few patient derived SBNET cell lines have been generated. Well-differentiated SBNET cells are slow growing and are hard to propagate. The few cell lines that have been established are not readily available, and after time in culture may not continue to express characteristics of NET cells. Generating new cell lines could take many years since SBNET cells have a long doubling time and many enrichment steps are needed in order to eliminate the rapidly dividing cancer-associated fibroblasts. To overcome these limitations, we have developed a protocol to culture SBNET cells from surgically removed tumors as spheroids in extracellular matrix (ECM). The ECM forms a 3-dimensional matrix that encapsulates SBNET cells and mimics the tumor micro-environment for allowing SBNET cells to grow. Here, we characterized the growth rate of SBNET spheroids and described methods to identify SBNET markers using immunofluorescence microscopy and immunohistochemistry to confirm that the spheroids are neuroendocrine tumor cells. In addition, we used SBNET spheroids for testing the cytotoxicity of rapamycin.

Immunohistochemistry

Enterochromaffin Cells - chemistry

Enterochromaffin Cells - pathology

Humans

Intestinal Neoplasms - chemistry

Intestinal Neoplasms - pathology

Intestine, Small - chemistry

Intestine, Small - pathology

Neuroendocrine Tumors - chemistry

Neuroendocrine Tumors - pathology

Pancreatic Neoplasms - chemistry

Pancreatic Neoplasms - pathology

Spheroids, Cellular - chemistry

Spheroids, Cellular - pathology

Stomach Neoplasms - chemistry

Stomach Neoplasms - pathology

Tumor Cells, Cultured

Tumor Microenvironment - physiology

Details

Title: Subtitle: Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids
Creators: Po Hien Ear - University of Iowa
Guiying Li - University of Iowa
Meng Wu - University of Iowa
Ellen Abusada - University of Iowa
Andrew M Bellizzi - University of Iowa
James R Howe - University of Iowa
Resource Type: Journal article
Publication Details: Journal of visualized experiments : JoVE, Vol.2019(152), e60303
DOI: 10.3791/60303
PMID: 31657801
PMCID: PMC7654522
NLM abbreviation: J Vis Exp
ISSN: 1940-087X
eISSN: 1940-087X
Grant note: R50 CA243786 / NCI NIH HHS P30 CA086862 / NCI NIH HHS P50 CA174521 / NCI NIH HHS
Language: English
Date published: 10/14/2019
Academic Unit: Pharmacy; Pathology; Surgery; Internal Medicine
Record Identifier: 9984183984502771

Metrics

14 Record Views

16 Times Cited - Web of Science