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Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients

Schwannomatosis, a rare form of neurofibromatosis, is characterized predominantly by multiple, often painful, schwannomas throughout the peripheral nervous system. The current standard of care for schwannomatosis is surgical resection. A major obstacle to schwannomatosis research is the lack of robu...

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Autores principales: Ostrow, Kimberly Laskie, Donaldson, Katelyn, Blakeley, Jaishri, Belzberg, Allan, Hoke, Ahmet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682832/
https://www.ncbi.nlm.nih.gov/pubmed/26657314
http://dx.doi.org/10.1371/journal.pone.0144620
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author Ostrow, Kimberly Laskie
Donaldson, Katelyn
Blakeley, Jaishri
Belzberg, Allan
Hoke, Ahmet
author_facet Ostrow, Kimberly Laskie
Donaldson, Katelyn
Blakeley, Jaishri
Belzberg, Allan
Hoke, Ahmet
author_sort Ostrow, Kimberly Laskie
collection PubMed
description Schwannomatosis, a rare form of neurofibromatosis, is characterized predominantly by multiple, often painful, schwannomas throughout the peripheral nervous system. The current standard of care for schwannomatosis is surgical resection. A major obstacle to schwannomatosis research is the lack of robust tumor cell lines. There is a great need for mechanistic and drug discovery studies of schwannomatosis, yet appropriate tools are not currently available. Schwannomatosis tumors are difficult to grow in culture as they survive only a few passages before senescence. Our lab has extensive experience in establishing primary and immortalized human Schwann cell cultures from normal tissue that retain their phenotypes after immortalization. Therefore we took on the challenge of creating immortalized human Schwann cell lines derived from tumors from schwannomatosis patients. We have established and fully characterized 2 schwannomatosis cell lines from 2 separate patients using SV40 virus large T antigen. One patient reported pain and the other did not. The schwannomatosis cell lines were stained with S100B antibodies to confirm Schwann cell identity. The schwannomatosis cells also expressed the Schwann cell markers, p75NTR, S100B, and NGF after multiple passages. Cell morphology was retained following multiple passaging and freeze/ thaw cycles. Gene expression microarray analysis was used to compare the cell lines with their respective parent tumors. No differences in key genes were detected, with the exception that several cell cycle regulators were upregulated in the schwannomatosis cell lines when compared to their parent tumors. This upregulation was apparently a product of cell culturing, as the schwannomatosis cells exhibited the same expression pattern of cell cycle regulatory genes as normal primary human Schwann cells. Cell growth was also similar between normal primary and immortalized tumor cells in culture. Accurate cell lines derived directly from human tumors will serve as invaluable tools for advancing schwannomatosis research, including drug screening.
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spelling pubmed-46828322015-12-31 Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients Ostrow, Kimberly Laskie Donaldson, Katelyn Blakeley, Jaishri Belzberg, Allan Hoke, Ahmet PLoS One Research Article Schwannomatosis, a rare form of neurofibromatosis, is characterized predominantly by multiple, often painful, schwannomas throughout the peripheral nervous system. The current standard of care for schwannomatosis is surgical resection. A major obstacle to schwannomatosis research is the lack of robust tumor cell lines. There is a great need for mechanistic and drug discovery studies of schwannomatosis, yet appropriate tools are not currently available. Schwannomatosis tumors are difficult to grow in culture as they survive only a few passages before senescence. Our lab has extensive experience in establishing primary and immortalized human Schwann cell cultures from normal tissue that retain their phenotypes after immortalization. Therefore we took on the challenge of creating immortalized human Schwann cell lines derived from tumors from schwannomatosis patients. We have established and fully characterized 2 schwannomatosis cell lines from 2 separate patients using SV40 virus large T antigen. One patient reported pain and the other did not. The schwannomatosis cell lines were stained with S100B antibodies to confirm Schwann cell identity. The schwannomatosis cells also expressed the Schwann cell markers, p75NTR, S100B, and NGF after multiple passages. Cell morphology was retained following multiple passaging and freeze/ thaw cycles. Gene expression microarray analysis was used to compare the cell lines with their respective parent tumors. No differences in key genes were detected, with the exception that several cell cycle regulators were upregulated in the schwannomatosis cell lines when compared to their parent tumors. This upregulation was apparently a product of cell culturing, as the schwannomatosis cells exhibited the same expression pattern of cell cycle regulatory genes as normal primary human Schwann cells. Cell growth was also similar between normal primary and immortalized tumor cells in culture. Accurate cell lines derived directly from human tumors will serve as invaluable tools for advancing schwannomatosis research, including drug screening. Public Library of Science 2015-12-14 /pmc/articles/PMC4682832/ /pubmed/26657314 http://dx.doi.org/10.1371/journal.pone.0144620 Text en © 2015 Ostrow et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Ostrow, Kimberly Laskie
Donaldson, Katelyn
Blakeley, Jaishri
Belzberg, Allan
Hoke, Ahmet
Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients
title Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients
title_full Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients
title_fullStr Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients
title_full_unstemmed Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients
title_short Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients
title_sort immortalized human schwann cell lines derived from tumors of schwannomatosis patients
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4682832/
https://www.ncbi.nlm.nih.gov/pubmed/26657314
http://dx.doi.org/10.1371/journal.pone.0144620
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