The Cell Morphology Core (CMC) supports the grant members with all aspects of histology-related work necessary to accomplish their research goals. The core is directed by Dr. Peter Bell and is housed in 800 square feet of dedicated space. It owns all the equipment necessary to process tissues and cells for a wide array of histological analyses. This includes several fluorescence microscopes, a transmission electron microscope, microtomes, an ultramicrotome, several cryostats, and automated tissue processing equipment. For tissue analysis at the light microscope level, paraffin, frozen and plastic-embedded samples are used. Routine and special stains as well as the detection of reporter gene expression in tissues (GFP, LacZ) are performed regularly. Further techniques include immunostaining, in situ hybridization, apoptosis and cell proliferation assays. Immunostaining is performed with fluorescent or enzyme-labeled antibodies, using all the appropriate positive and negative controls. In situ hybridization for identification of transduced cells or the detection of viruses uses labeled DMA or RNA probes. Images can be acquired by widefield or confocal microscopy and analyzed with Image-Pro Plus and AnalySIS software, quantifying the degree of staining for any target. Transmission electron microscopy (TEM) is performed to evaluate tissue samples at an ultrastructural level. For vector analysis, negative staining of viral preparations is applied for EM analysis. Viral vectors are assessed for average density, purity, uniformity and viral particle integrity. Contaminations in adenovirus preparations with AAV (and vice versa) are detectable due to the difference in size of both viruses. Investigators can also receive training in histological techniques and microscopy allowing them to perform their own analyses by utilizing the core's resources.

Public Health Relevance

The Cell Morphology core processes and analyzes tissues both for histopathology and the expression of transgenes, important parameters to assess the success of gene therapy studies. The histological examination of tissues is aimed to detect how gene transfer experiments affect target tissues and other organs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Center Core Grants (P30)
Project #
5P30DK047757-20
Application #
8448746
Study Section
Special Emphasis Panel (ZDK1-GRB-1)
Project Start
Project End
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
20
Fiscal Year
2013
Total Cost
$77,946
Indirect Cost
$28,396
Name
University of Pennsylvania
Department
Type
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Gurda, Brittney L; De Guilhem De Lataillade, Adrien; Bell, Peter et al. (2016) Evaluation of AAV-mediated Gene Therapy for Central Nervous System Disease in Canine Mucopolysaccharidosis VII. Mol Ther 24:206-16
Ortved, Kyla; Wagner, Bettina; Calcedo, Roberto et al. (2015) Humoral and cell-mediated immune response, and growth factor synthesis after direct intraarticular injection of rAAV2-IGF-I and rAAV5-IGF-I in the equine middle carpal joint. Hum Gene Ther 26:161-71
Bissig-Choisat, Beatrice; Wang, Lili; Legras, Xavier et al. (2015) Development and rescue of human familial hypercholesterolaemia in a xenograft mouse model. Nat Commun 6:7339
Wang, Lili; Bell, Peter; Somanathan, Suryanarayan et al. (2015) Comparative Study of Liver Gene Transfer With AAV Vectors Based on Natural and Engineered AAV Capsids. Mol Ther 23:1877-87
Calcedo, Roberto; Franco, Judith; Qin, Qiuyue et al. (2015) Preexisting Neutralizing Antibodies to Adeno-Associated Virus Capsids in Large Animals Other Than Monkeys May Confound In Vivo Gene Therapy Studies. Hum Gene Ther Methods 26:103-5
Baek, Jeong-In; Choi, Soo Young; Chacon-Heszele, Maria F et al. (2014) Expression of Drosophila forkhead transcription factors during kidney development. Biochem Biophys Res Commun 446:15-7
O'Neill, S M; Hinkle, C; Chen, S-J et al. (2014) Targeting adipose tissue via systemic gene therapy. Gene Ther 21:653-61
Huang, Liwei; Xiao, An; Wecker, Andrea et al. (2014) A possible zebrafish model of polycystic kidney disease: knockdown of wnt5a causes cysts in zebrafish kidneys. J Vis Exp :
Huang, Liwei; Xiao, An; Choi, Soo Young et al. (2014) Wnt5a is necessary for normal kidney development in zebrafish and mice. Nephron Exp Nephrol 128:80-8
Adam, Virginie S; Crosariol, Marco; Kumar, Sachin et al. (2014) Adeno-associated virus 9-mediated airway expression of antibody protects old and immunodeficient mice against influenza virus. Clin Vaccine Immunol 21:1528-33

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