Methodology, Criteria, and Characterization of Patient-Matched Thyroid Cell Lines and Patient-Derived Tumor Xenografts

Laura A. Marlow, Stephen D. Rohl, James L. Miller, Jeffery A. Knauf, James A. Fagin, Mabel Ryder, Dragana Milosevic, Brian C. Netzel, Stefan K. Grebe, Honey V. Reddi, Robert Christian Smallridge, John A III Copland

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Objective: To investigate the molecular underpinnings of thyroid cancer, preclinical cell line models are crucial; however, ;40% of these have been proven to be either duplicates of existing thyroid lines or even nonthyroid-derived lines or are not derived from humans at all. Therefore, we set out to establish procedures and guidelines that should proactively avoid these problems, which facilitated the creation of criteria to make valid preclinical models for thyroid cancer research. Design: Based on our recommendations, we systematically characterized all new cell lines that we generated by a standardized approach that included (1) determination of human origin, (2) exclusion of lymphoma, (3) DNA fingerprinting and histological comparisons to establish linkage to presumed tissue of origin, (4) examining thyroid differentiation by screening two to three thyroid markers, (5) examination of biological behavior (growth rate, tumorigenicity), and (6) presence of common thyroid cancer genetic changes (TP53, BRAF, PTEN, PIK3CA, RAS, TERT promoter, RET/PTC, PAX8/PPARg, NF1, and EIF1AX). Results: We established seven new thyroid cell lines (LAM136, EAM306, SDAR1, SDAR2, JEM493, THJ529, and THJ560) out of 294 primary culture attempts, and 10 patient-derived tumor xenografts (PDTXs; MC-Th-95, MC-Th-374, MC-Th-467, MC-Th-491, MC-Th-493, MC-Th-504, MC-Th-524, MC-Th- 529, MC-Th-560, and MC-Th-562) out of 67 attempts. All were successfully validated by our protocols. Conclusions: This standardized approach for cell line and PDTX characterization should prevent (or detect) future cross-contamination and ensure that only valid preclinical models are used for thyroid cancer research.

Original languageEnglish (US)
Pages (from-to)3169-3182
Number of pages14
JournalJournal of Clinical Endocrinology and Metabolism
Volume103
Issue number9
DOIs
StatePublished - Sep 1 2018

Fingerprint

Thyroid Neoplasms
Heterografts
Tumors
Thyroid Gland
Cells
Cell Line
Neoplasms
Factor IX
DNA Fingerprinting
Research
Lymphoma
Screening
Contamination
Guidelines
Tissue
DNA
Growth

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Methodology, Criteria, and Characterization of Patient-Matched Thyroid Cell Lines and Patient-Derived Tumor Xenografts. / Marlow, Laura A.; Rohl, Stephen D.; Miller, James L.; Knauf, Jeffery A.; Fagin, James A.; Ryder, Mabel; Milosevic, Dragana; Netzel, Brian C.; Grebe, Stefan K.; Reddi, Honey V.; Smallridge, Robert Christian; Copland, John A III.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 103, No. 9, 01.09.2018, p. 3169-3182.

Research output: Contribution to journalArticle

Marlow, Laura A. ; Rohl, Stephen D. ; Miller, James L. ; Knauf, Jeffery A. ; Fagin, James A. ; Ryder, Mabel ; Milosevic, Dragana ; Netzel, Brian C. ; Grebe, Stefan K. ; Reddi, Honey V. ; Smallridge, Robert Christian ; Copland, John A III. / Methodology, Criteria, and Characterization of Patient-Matched Thyroid Cell Lines and Patient-Derived Tumor Xenografts. In: Journal of Clinical Endocrinology and Metabolism. 2018 ; Vol. 103, No. 9. pp. 3169-3182.
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T1 - Methodology, Criteria, and Characterization of Patient-Matched Thyroid Cell Lines and Patient-Derived Tumor Xenografts

AU - Marlow, Laura A.

AU - Rohl, Stephen D.

AU - Miller, James L.

AU - Knauf, Jeffery A.

AU - Fagin, James A.

AU - Ryder, Mabel

AU - Milosevic, Dragana

AU - Netzel, Brian C.

AU - Grebe, Stefan K.

AU - Reddi, Honey V.

AU - Smallridge, Robert Christian

AU - Copland, John A III

PY - 2018/9/1

Y1 - 2018/9/1

N2 - Objective: To investigate the molecular underpinnings of thyroid cancer, preclinical cell line models are crucial; however, ;40% of these have been proven to be either duplicates of existing thyroid lines or even nonthyroid-derived lines or are not derived from humans at all. Therefore, we set out to establish procedures and guidelines that should proactively avoid these problems, which facilitated the creation of criteria to make valid preclinical models for thyroid cancer research. Design: Based on our recommendations, we systematically characterized all new cell lines that we generated by a standardized approach that included (1) determination of human origin, (2) exclusion of lymphoma, (3) DNA fingerprinting and histological comparisons to establish linkage to presumed tissue of origin, (4) examining thyroid differentiation by screening two to three thyroid markers, (5) examination of biological behavior (growth rate, tumorigenicity), and (6) presence of common thyroid cancer genetic changes (TP53, BRAF, PTEN, PIK3CA, RAS, TERT promoter, RET/PTC, PAX8/PPARg, NF1, and EIF1AX). Results: We established seven new thyroid cell lines (LAM136, EAM306, SDAR1, SDAR2, JEM493, THJ529, and THJ560) out of 294 primary culture attempts, and 10 patient-derived tumor xenografts (PDTXs; MC-Th-95, MC-Th-374, MC-Th-467, MC-Th-491, MC-Th-493, MC-Th-504, MC-Th-524, MC-Th- 529, MC-Th-560, and MC-Th-562) out of 67 attempts. All were successfully validated by our protocols. Conclusions: This standardized approach for cell line and PDTX characterization should prevent (or detect) future cross-contamination and ensure that only valid preclinical models are used for thyroid cancer research.

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