Nod cb17 prkdcscid j female mice

SKNMM^MYCN and SKNMM^CONT cells were labeled with luciferase-YFP (yellow fluorescent protein) using a lentivirus. Two mixed cell pools were prepared in a 1:1 ratio: labeled SKNMM^MYCN cells/unlabeled SKNMM^CONT and labeled SKNMM^CONT /unlabeled SKNMM^MYCN. Cell mixes were injected in the para-adrenal region of the mice (four million cells/mouse) with ultrasound guidance under anesthesia as explained before. In this experiment, we used NOD.CB17-Prkdc^scid/J female mice (Jackson laboratories, Stock # 001303). The mice were followed up for the tumor formation using ultrasound scanning every 2 weeks, xenogen signal every week and palpation every week. Mice were enrolled in the study when the tumor was at least 14 mm³ in size using the ultrasound scanning. Enrolled mice were randomly assigned to receive either doxycycline (2 mg/ml) in the drinking water or continue on regular water.
The labeled/unlabeled cell mixes were also maintained in culture and the ratio of YFP-expressing cells in each cell mix was determined weekly using flow cytometric analysis.

Animal protocols and experimental procedures are approved by the Institutional Animal Care and Use Committee at San Diego State University. A total of 10 NOD.CB17-Prkdc^scid/J female mice (The Jackson Laboratory, catalog #001303) under 2% isoflurane (Victor Medical, catalog #NDC 57319-474-06) each received three intramyocardial injections of CardioClusters + FluoSpheres tracking beads (Thermo Fisher Scientific, catalog #F8815; 1:50,000 dilution) in LV wall. A total of 90,000 cells per heart were introduced into CardioCluster animals. The hearts were immediately placed back into the intrathoracic space followed by muscle and skin closure. All animals were euthanized 3-days post injection for histological examination of injection location.

All animal experiments were approved by and performed in accordance with the guidelines of the University of Miami Institutional Animal Care and Use Committee. For live visualization of Apt63 tumor uptake and retention in vivo, a prostate xenograft tumor model was used. NOD.CB17-Prkdcscid/J male mice (10 weeks old, n = 14, The Jackson Laboratory) were injected orthotopically into the right anterior lobe of the prostate with 2 × 10⁶ LN3 and 2 × 10⁶ Pro5 cells. For Apt63 cytotoxicity in vivo, we used a previously described breast tumor xenograft mouse model [24 (link)]. NOD.CB17-Prkdcscid/J female mice (6–8 weeks old, n = 39, The Jackson Laboratory) were injected with 10⁶ of MDA-MB-231 cells into the mammary fat pad. In both xenograft models, when tumors were palpable, 1 nmol of Alexa Fluor™ 647-labeled Apt63, AptScr, or unlabeled oligonucleotides suspended in 200 µL of PBS was injected into the tail vein as a single injection. Mice were imaged and euthanized at specified time points. Tumors and selected tissues were dissected and processed for further analysis. Detailed procedures are described in Online Resource 2.