Detection and measurement of in vitro gene transfer by gamma camera imaging

K. R. Zinn, T. R. Chaudhuri, D. J. Buchsbaum, J. M. Mountz, B. E. Rogers

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

The purpose of this work was to develop a high capacity method to image gene transfer to cancer cells growing as monolayers in cell culture plates. A sensitive and high capacity nuclear-imaging method for detection of gene transfer in vitro will allow rapid validation of vectors in different cell lines under various conditions. Human cancer cell lines (A-427 non-small cell lung, SKOV3.ip1 ovarian, MDA-MB-468 breast, and BxPC-3 pancreatic) were infected with a replication-incompetent adenoviral vector encoding the human type 2 somatostatin receptor (Ad-hSSTr2). Expression of the hSSTr2 reporter protein in cells was detected by imaging an internalized 99mTc-labeled, hSSTr2 binding peptide (P2045, Diatide, Inc.). Imaging provided an accurate measure of internally bound 99mTc as evidenced by equivalence of results for imaging region of interest (ROI) analyses and gamma counter measurements. Internally bound 99mTc-P2045 was linearly correlated (R2 = 0.98) with the percentage of hSSTr2-positive cells following gene transfer. Excess P2045 blocked binding and internalization of the 99mTc-P2045, indicating the specificity of the technique. Up to four 96-well plates could be imaged simultaneously, thereby demonstrating the high capacity of the system. This novel in vitro approach provides a new method to test enhanced gene transfer as new vectors are developed.

Original languageEnglish
Pages (from-to)291-299
Number of pages9
JournalGene therapy
Volume8
Issue number4
DOIs
StatePublished - Apr 5 2001
Externally publishedYes

Keywords

  • Gene transfer
  • Imaging
  • Somatostatin receptor
  • Tc

Fingerprint Dive into the research topics of 'Detection and measurement of in vitro gene transfer by gamma camera imaging'. Together they form a unique fingerprint.

  • Cite this