Isostructural fluorescent and radioactive probes for monitoring neural stem and progenitor cell transplants.
A construct for tagging neurospheres and monitoring cell transplantations was developed using a new technology for producing luminescent and radiolabeled probes that have identical structures. The HIV1-Tat basic domain derivatives NAcGRKKRRQRRR(SAACQ)G (SAACQ-1) and [NAcGRKKRRQRRR(Re(CO)3SAACQ)G]+ (ReSAACQ-1) were prepared in excellent yields using the single amino acid chelate-quinoline (SAACQ) ligand and its Re(I) complex and conventional automated peptide synthesis methods. The distribution of the luminescent Re probe, using epifluorescence microscopy, showed that it localized primarily in the cell nucleus with a significant degree of association on the nuclear envelope. A smaller amount was found to be dispersed in the cytoplasm. The 99m Tc analogue was then prepared in 43+/-7% (n=12) yield and very high effective specific activity. Following incubation, average uptake of the probe in neurospheres ranged between 10 and 20 Bq/cell. As determined by colorimetric assays, viability for cells labeled with high effective specific activity 99m TcSAACQ-1 was 97+/-4% at 2 h postlabeling and 85+/-25% at 24 h postlabeling for incubation activities ranging from 245 to 8900 Bq/cell. DNA analysis showed that at these levels, there was no significant difference between the extent of DNA damage in the treated cells versus control cells. A series of preliminary SPECT/CT studies of transplants in mice were performed, which showed that the strategy is convenient and feasible and that it is possible to routinely assess procedures noninvasively and determine the number of cells transplanted.
Schaffer
Paul
P
McMaster Nuclear Reactor, McMaster University, Hamilton, Ontario, Canada L8S 4K1.
Gleave
Jacqueline A
JA
Lemon
Jennifer A
JA
Reid
Leslie C
LC
Pacey
Laura K K
LK
Farncombe
Troy H
TH
Boreham
Douglas R
DR
Zubieta
Jon
J
Babich
John W
JW
Doering
Laurie C
LC
Valliant
John F
JF
D000818:Animals; D002478:Cells, Cultured; D002614:Chelating Agents / Q000737:chemistry; D003593:Cytoplasm / Q000000981:diagnostic imaging / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006257:Head / Q000000981:diagnostic imaging / Q000473:pathology; D007553:Isotope Labeling; D008164:Luminescent Proteins / Q000138:chemical synthesis* / Q000493:pharmacokinetics*; D051379:Mice; D009420:Nervous System / Q000166:cytology / Q000000981:diagnostic imaging; D009685:Nuclear Envelope / Q000000981:diagnostic imaging / Q000378:metabolism; D011804:Quinolines / Q000737:chemistry; D019275:Radiopharmaceuticals / Q000138:chemical synthesis* / Q000493:pharmacokinetics*; D013194:Staining and Labeling / Q000379:methods; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000000981:diagnostic imaging; D013667:Technetium; D015899:Tomography, Emission-Computed, Single-Photon; D014057:Tomography, X-Ray Computed; D051598:Whole Body Imaging; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000493:pharmacokinetics
D002614:Chelating Agents; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011804:Quinolines; D019275:Radiopharmaceuticals; D054322:tat Gene Products, Human Immunodeficiency Virus; D013667:Technetium; C037219:quinoline