In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue

V. K. Verma, S. S. Beevi, A. Tabassum, K. Kumaresan, R. S. Kamaraju, A. S. Arbab, L. K. Chelluri

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Introduction: Noninvasive radionuclide imaging of cells using technetium99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) is a potential diagnostic tool for several applications. Herein we aimed to evaluate the labeling efficiency and cellular toxicity of 99mTc-HMPAO with Stromal Vascular Fraction (SVF) of adipose tissue to develop a process tool for theranostic purposes, in particular imaging cardiac stem cell therapy. Methods: Ten million cells of SVF were labeled with 99mTc-HMPAO complex and excess radiolabel was cleared off through washing in PBS. The labeling efficiency of 99mTc-HMPAO was detected in labeled cells and their subsequent supernatant wash using isotope dose calibrator and gamma camera. The cytotoxicity was assessed for the comparative reactive oxygen species (ROS) by H2DCFDDA, apoptotic events by annexin-V and TUNEL assay and mitochondrial potential by JC-1. Results: An encouraging labeling efficiency of 33% was observed with 99mTc-HMPAO complex. The radionuclide labeling of SVF demonstrated significant safety profile as evaluated by apoptotic assays. Conclusion: 99mTc-HMPAO labeling efficiency of 33% of total SV fraction would produce sufficient radioactive signals that would enable for in vivo tracking of cells by SPECT-CT. The radionuclide did not demonstrate any significant impact on the structural or functional organization of the labeled cells. Our study indicates that SVF can be safely labeled with 99mTc-HMPAO without adverse cytotoxic events and for its potential role in imaging cardiac stem cell therapy.

Original languageEnglish (US)
Pages (from-to)744-748
Number of pages5
JournalNuclear Medicine and Biology
Volume41
Issue number9
DOIs
StatePublished - Jan 1 2014

Fingerprint

Technetium Tc 99m Exametazime
Blood Vessels
Adipose Tissue
Cell- and Tissue-Based Therapy
Radioisotopes
Stem Cells
Cell Tracking
Gamma Cameras
Annexin A5
In Situ Nick-End Labeling
Stromal Cells
In Vitro Techniques
Isotopes
Radionuclide Imaging
Reactive Oxygen Species
Safety

Keywords

  • Cardiac stem cell therapy
  • Cytotoxicity
  • Labeling efficiency
  • SVF
  • Tc-HMPAO

ASJC Scopus subject areas

  • Molecular Medicine
  • Radiology Nuclear Medicine and imaging
  • Cancer Research

Cite this

In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue. / Verma, V. K.; Beevi, S. S.; Tabassum, A.; Kumaresan, K.; Kamaraju, R. S.; Arbab, A. S.; Chelluri, L. K.

In: Nuclear Medicine and Biology, Vol. 41, No. 9, 01.01.2014, p. 744-748.

Research output: Contribution to journalArticle

Verma, V. K. ; Beevi, S. S. ; Tabassum, A. ; Kumaresan, K. ; Kamaraju, R. S. ; Arbab, A. S. ; Chelluri, L. K. / In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue. In: Nuclear Medicine and Biology. 2014 ; Vol. 41, No. 9. pp. 744-748.
@article{99b0fbf5169e405cbc8c67e355dd0ce2,
title = "In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue",
abstract = "Introduction: Noninvasive radionuclide imaging of cells using technetium99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) is a potential diagnostic tool for several applications. Herein we aimed to evaluate the labeling efficiency and cellular toxicity of 99mTc-HMPAO with Stromal Vascular Fraction (SVF) of adipose tissue to develop a process tool for theranostic purposes, in particular imaging cardiac stem cell therapy. Methods: Ten million cells of SVF were labeled with 99mTc-HMPAO complex and excess radiolabel was cleared off through washing in PBS. The labeling efficiency of 99mTc-HMPAO was detected in labeled cells and their subsequent supernatant wash using isotope dose calibrator and gamma camera. The cytotoxicity was assessed for the comparative reactive oxygen species (ROS) by H2DCFDDA, apoptotic events by annexin-V and TUNEL assay and mitochondrial potential by JC-1. Results: An encouraging labeling efficiency of 33{\%} was observed with 99mTc-HMPAO complex. The radionuclide labeling of SVF demonstrated significant safety profile as evaluated by apoptotic assays. Conclusion: 99mTc-HMPAO labeling efficiency of 33{\%} of total SV fraction would produce sufficient radioactive signals that would enable for in vivo tracking of cells by SPECT-CT. The radionuclide did not demonstrate any significant impact on the structural or functional organization of the labeled cells. Our study indicates that SVF can be safely labeled with 99mTc-HMPAO without adverse cytotoxic events and for its potential role in imaging cardiac stem cell therapy.",
keywords = "Cardiac stem cell therapy, Cytotoxicity, Labeling efficiency, SVF, Tc-HMPAO",
author = "Verma, {V. K.} and Beevi, {S. S.} and A. Tabassum and K. Kumaresan and Kamaraju, {R. S.} and Arbab, {A. S.} and Chelluri, {L. K.}",
year = "2014",
month = "1",
day = "1",
doi = "10.1016/j.nucmedbio.2014.06.002",
language = "English (US)",
volume = "41",
pages = "744--748",
journal = "Nuclear Medicine and Biology",
issn = "0969-8051",
publisher = "Elsevier Inc.",
number = "9",

}

TY - JOUR

T1 - In vitro assessment of cytotoxicity and labeling efficiency of 99mTc-HMPAO with stromal vascular fraction of adipose tissue

AU - Verma, V. K.

AU - Beevi, S. S.

AU - Tabassum, A.

AU - Kumaresan, K.

AU - Kamaraju, R. S.

AU - Arbab, A. S.

AU - Chelluri, L. K.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Introduction: Noninvasive radionuclide imaging of cells using technetium99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) is a potential diagnostic tool for several applications. Herein we aimed to evaluate the labeling efficiency and cellular toxicity of 99mTc-HMPAO with Stromal Vascular Fraction (SVF) of adipose tissue to develop a process tool for theranostic purposes, in particular imaging cardiac stem cell therapy. Methods: Ten million cells of SVF were labeled with 99mTc-HMPAO complex and excess radiolabel was cleared off through washing in PBS. The labeling efficiency of 99mTc-HMPAO was detected in labeled cells and their subsequent supernatant wash using isotope dose calibrator and gamma camera. The cytotoxicity was assessed for the comparative reactive oxygen species (ROS) by H2DCFDDA, apoptotic events by annexin-V and TUNEL assay and mitochondrial potential by JC-1. Results: An encouraging labeling efficiency of 33% was observed with 99mTc-HMPAO complex. The radionuclide labeling of SVF demonstrated significant safety profile as evaluated by apoptotic assays. Conclusion: 99mTc-HMPAO labeling efficiency of 33% of total SV fraction would produce sufficient radioactive signals that would enable for in vivo tracking of cells by SPECT-CT. The radionuclide did not demonstrate any significant impact on the structural or functional organization of the labeled cells. Our study indicates that SVF can be safely labeled with 99mTc-HMPAO without adverse cytotoxic events and for its potential role in imaging cardiac stem cell therapy.

AB - Introduction: Noninvasive radionuclide imaging of cells using technetium99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) is a potential diagnostic tool for several applications. Herein we aimed to evaluate the labeling efficiency and cellular toxicity of 99mTc-HMPAO with Stromal Vascular Fraction (SVF) of adipose tissue to develop a process tool for theranostic purposes, in particular imaging cardiac stem cell therapy. Methods: Ten million cells of SVF were labeled with 99mTc-HMPAO complex and excess radiolabel was cleared off through washing in PBS. The labeling efficiency of 99mTc-HMPAO was detected in labeled cells and their subsequent supernatant wash using isotope dose calibrator and gamma camera. The cytotoxicity was assessed for the comparative reactive oxygen species (ROS) by H2DCFDDA, apoptotic events by annexin-V and TUNEL assay and mitochondrial potential by JC-1. Results: An encouraging labeling efficiency of 33% was observed with 99mTc-HMPAO complex. The radionuclide labeling of SVF demonstrated significant safety profile as evaluated by apoptotic assays. Conclusion: 99mTc-HMPAO labeling efficiency of 33% of total SV fraction would produce sufficient radioactive signals that would enable for in vivo tracking of cells by SPECT-CT. The radionuclide did not demonstrate any significant impact on the structural or functional organization of the labeled cells. Our study indicates that SVF can be safely labeled with 99mTc-HMPAO without adverse cytotoxic events and for its potential role in imaging cardiac stem cell therapy.

KW - Cardiac stem cell therapy

KW - Cytotoxicity

KW - Labeling efficiency

KW - SVF

KW - Tc-HMPAO

UR - http://www.scopus.com/inward/record.url?scp=84926259316&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84926259316&partnerID=8YFLogxK

U2 - 10.1016/j.nucmedbio.2014.06.002

DO - 10.1016/j.nucmedbio.2014.06.002

M3 - Article

C2 - 25037755

AN - SCOPUS:84926259316

VL - 41

SP - 744

EP - 748

JO - Nuclear Medicine and Biology

JF - Nuclear Medicine and Biology

SN - 0969-8051

IS - 9

ER -