Fluorescent detection of Merlin-deficient Schwann cells and primary human vestibular schwannoma cells using sodium fluorescein

Enrique R. Perez, Olena Bracho, Liliana Ein, Mikhaylo Szczupak, Paula V Monje, Cristina Fernandez-Valle, Abdulaziz Alshaiji, Michael Ivan, Jacques Morcos, Xue Z Liu, Michael E Hoffer, Adrien Eshraghi, Simon I Angeli, Fred F Telischi, Christine T Dinh

Research output: Contribution to journalArticle

Abstract

Hypothesis: Merlin-deficient Schwann cells (MD-SC) and primary human vestibular schwannoma (VS) cells exhibit selective uptake of sodium-fluorescein (SF), allowing for fluorescent detection and improved visualization of tumor cells, when compared with Schwann cells (SC). Background: SF is a fluorescent compound used for fluorescence- guided resection of gliomas. The utility of SF for VS surgery has not been assessed. Methods: Mouse MD-SCs and rat SCs were cultured on 96- well plates at different cell densities and treated with SF at several drug concentrations and durations. Relative fluorescence units (RFU) were measured using a fluorometer to determine optimal treatment parameters in vitro. Subsequently, a four-point Likert scale for fluorescence visualization of pelleted cells was created and validated. Blinded observers rated SF-treated primary human VS and SC cultures, which were developed from deidentified specimens obtained from live and cadaveric donors, respectively. Results: In contrast to SCs that showed low levels of fluorescence, MD-SCs demonstrated dose-dependent increases in RFUs when treated with incremental dosages of SF as well as longer treatment and fluorescent excitation times. In addition, RFUs were higher at greater MD-SC densities. The Likert scale for fluorescence visualization was validated using nine blinded observers and there were excellent inter- and intrarater reliabilities (intraclass coefficients of 0.989 and >0.858, respectively). Using the Likert scale, human VS treated with SF received higher scores than human SCs ( p<0.001). Conclusion: Mouse MD-SC and human VS cells demonstrate preferential uptake of SF when compared with normal primary SCs. Observers detected differences in fluorescence using the validated Likert scale. Further investigations into the utility of SF-guidance in VS surgery are warranted.

Original languageEnglish (US)
Pages (from-to)1053-1059
Number of pages7
JournalOtology and Neurotology
Volume39
Issue number8
DOIs
StatePublished - Jan 1 2018

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Neurofibromin 2
Acoustic Neuroma
Schwann Cells
Fluorescein
Fluorescence
Cell Count
Glioma
Cell Culture Techniques

Keywords

  • Fluorescein
  • Fluorescence
  • Schwann
  • Schwannoma
  • Sodium fluorescein
  • Vestibular schwannoma
  • VS

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Sensory Systems
  • Clinical Neurology

Cite this

Fluorescent detection of Merlin-deficient Schwann cells and primary human vestibular schwannoma cells using sodium fluorescein. / Perez, Enrique R.; Bracho, Olena; Ein, Liliana; Szczupak, Mikhaylo; Monje, Paula V; Fernandez-Valle, Cristina; Alshaiji, Abdulaziz; Ivan, Michael; Morcos, Jacques; Liu, Xue Z; Hoffer, Michael E; Eshraghi, Adrien; Angeli, Simon I; Telischi, Fred F; Dinh, Christine T.

In: Otology and Neurotology, Vol. 39, No. 8, 01.01.2018, p. 1053-1059.

Research output: Contribution to journalArticle

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abstract = "Hypothesis: Merlin-deficient Schwann cells (MD-SC) and primary human vestibular schwannoma (VS) cells exhibit selective uptake of sodium-fluorescein (SF), allowing for fluorescent detection and improved visualization of tumor cells, when compared with Schwann cells (SC). Background: SF is a fluorescent compound used for fluorescence- guided resection of gliomas. The utility of SF for VS surgery has not been assessed. Methods: Mouse MD-SCs and rat SCs were cultured on 96- well plates at different cell densities and treated with SF at several drug concentrations and durations. Relative fluorescence units (RFU) were measured using a fluorometer to determine optimal treatment parameters in vitro. Subsequently, a four-point Likert scale for fluorescence visualization of pelleted cells was created and validated. Blinded observers rated SF-treated primary human VS and SC cultures, which were developed from deidentified specimens obtained from live and cadaveric donors, respectively. Results: In contrast to SCs that showed low levels of fluorescence, MD-SCs demonstrated dose-dependent increases in RFUs when treated with incremental dosages of SF as well as longer treatment and fluorescent excitation times. In addition, RFUs were higher at greater MD-SC densities. The Likert scale for fluorescence visualization was validated using nine blinded observers and there were excellent inter- and intrarater reliabilities (intraclass coefficients of 0.989 and >0.858, respectively). Using the Likert scale, human VS treated with SF received higher scores than human SCs ( p<0.001). Conclusion: Mouse MD-SC and human VS cells demonstrate preferential uptake of SF when compared with normal primary SCs. Observers detected differences in fluorescence using the validated Likert scale. Further investigations into the utility of SF-guidance in VS surgery are warranted.",
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T1 - Fluorescent detection of Merlin-deficient Schwann cells and primary human vestibular schwannoma cells using sodium fluorescein

AU - Perez, Enrique R.

AU - Bracho, Olena

AU - Ein, Liliana

AU - Szczupak, Mikhaylo

AU - Monje, Paula V

AU - Fernandez-Valle, Cristina

AU - Alshaiji, Abdulaziz

AU - Ivan, Michael

AU - Morcos, Jacques

AU - Liu, Xue Z

AU - Hoffer, Michael E

AU - Eshraghi, Adrien

AU - Angeli, Simon I

AU - Telischi, Fred F

AU - Dinh, Christine T

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AB - Hypothesis: Merlin-deficient Schwann cells (MD-SC) and primary human vestibular schwannoma (VS) cells exhibit selective uptake of sodium-fluorescein (SF), allowing for fluorescent detection and improved visualization of tumor cells, when compared with Schwann cells (SC). Background: SF is a fluorescent compound used for fluorescence- guided resection of gliomas. The utility of SF for VS surgery has not been assessed. Methods: Mouse MD-SCs and rat SCs were cultured on 96- well plates at different cell densities and treated with SF at several drug concentrations and durations. Relative fluorescence units (RFU) were measured using a fluorometer to determine optimal treatment parameters in vitro. Subsequently, a four-point Likert scale for fluorescence visualization of pelleted cells was created and validated. Blinded observers rated SF-treated primary human VS and SC cultures, which were developed from deidentified specimens obtained from live and cadaveric donors, respectively. Results: In contrast to SCs that showed low levels of fluorescence, MD-SCs demonstrated dose-dependent increases in RFUs when treated with incremental dosages of SF as well as longer treatment and fluorescent excitation times. In addition, RFUs were higher at greater MD-SC densities. The Likert scale for fluorescence visualization was validated using nine blinded observers and there were excellent inter- and intrarater reliabilities (intraclass coefficients of 0.989 and >0.858, respectively). Using the Likert scale, human VS treated with SF received higher scores than human SCs ( p<0.001). Conclusion: Mouse MD-SC and human VS cells demonstrate preferential uptake of SF when compared with normal primary SCs. Observers detected differences in fluorescence using the validated Likert scale. Further investigations into the utility of SF-guidance in VS surgery are warranted.

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KW - Sodium fluorescein

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