More than 50 of the assessable animals in a group have grown > 자유게시판

More than 50 of the assessable animals in a group have grown to the endpoint volume or treatment exceeded the MTD (20 body weight loss or >10 TR related mortality).Serum lipaseCambridge, MA)]; BCL-10 [1:200, anti-rabbit] and CD31 [1:100, anti-goat (Santa Cruz, CA)]; BRCA2 [1:500, anti-mouse (R D systems)]. For IHC the Envision Dual Labeled Polymer kit (DakoCytomation) was used according to the manufacturer's instructions and then lightly counterstained with Gill I hematoxylin (Sigma-Aldrich) before dehydration and mounting. Images were obtained using Scanscope XT (Aperio Staurosporine Technologies, Vista, CA) and the staining of the TMA punches were scored using an algorithm in the Imagescope software (Aperio Technologies) created by a histologist based upon signal intensity (0, 1+ , 2+ , 3+) and percentage. For immunofluorescence, tissue was incubated with AlexaFlour 594-conjugated secondary antibody (Invitrogen) and mounted with Vectashield containing DAPI (Vector Laboratories, Burlingame, CA). Images were acquired on a Zeiss LSM 880 confocal laser scanning microscope (Carl Zeiss MicroImaging, Inc, Thornwood, NY).Gene mutation analysisDNA from a fresh frozen sample of PA-018 was isolated and underwent custom capture (eArray; Agilent, Santa Clara, CA) of the coding region and intron/exon boundaries of coding exons for BRCA2. Products from each capture reaction were sequenced on a HiSeq 2000 (Illumina, San Diego, CA) and analyzed as described by Couch et al. [21].ResultsCharacterization of the PACC PDTX mouse modelBlood was collected sublingually under no anesthesia at pre-dose and day 15 for n = 5 mice per group in EDTA collection tubes. Serum was collected and analyzed by IDEXX laboratories (Westbrook, Maine) for serum lipase levels.Immunohistochemistry (IHC) and immunofluorescence (IF)Formalin-fixed tissues were collected and embedded into paraffin. A tissue microarray (TMA) was constructed for all tumor tissues from each of the treatment groups and used for IHC analysis. TMA tissues were cut into 5 mm sections, deparaffinized, hydrated, antigen retrieved and blocked with diluent that contained Background Reducing Components (Dakocytomation, Denmark). Immunostaining was done on either the TMA or PA-018 alone with the following: carcinoembryonic antigen (CEA), neuron specific enolase (NSE), Chromogranin A (CgA), and cytokeratin 19 (CK19) [1:100, anti-mouse with rodent block (Dakocytomation)]; Mist-1 [1:2000] and cleaved caspase-3 (CC3) [1:100 hi pH, anti-rabbit (Cell Signaling, Beverly, MA)]; amylase [1:1000, anti-rabbit (Sigma-Aldrich, St.Louis, MO)]; lipase [1:1600], BRCA1 [1:100], and Collagen I [1:1500, anti-rabbit (Abcam,PA-018, a pancreatic acinar cell carcinoma (PACC) PDTX mouse model, was derived from a metastatic liver biopsy of a patient previously described in a case report [20]. The histologic features of PA-018 were similar to those in the patient's pancreatic primary tissue (Fig. 1a). These features included a solid, acinar-like growth pattern by cells with undifferentiated nuclei and amphophilic cytoplasm. Immunohistochemistry (IHC) with human specific mitochondrial surface protein and human lamin A+C and confirmed that the xenograft tumor cells were PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/17139194 of human origin (Fig. 1b). Short tandem repeat (STR) analysis showed that the genetic signature of our PDTX tumor (passage 5) closely matched (allele drop out was noted) the signature of the patient. The DNA that was used for comparison was from a sample taken 3 years prior to.