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
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
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
Xiao, Yan; Nagai, Yasuhiro; Deng, Guoping et al. (2014) Dynamic interactions between TIP60 and p300 regulate FOXP3 function through a structural switch defined by a single lysine on TIP60. Cell Rep 7:1471-80
Chacon-Heszele, Maria F; Zuo, Xiaofeng; Hellman, Nathan E et al. (2014) Novel MAPK-dependent and -independent tubulogenes identified via microarray analysis of 3D-cultured Madin-Darby canine kidney cells. Am J Physiol Renal Physiol 306:F1047-58
Mays, Lauren E; Wang, Lili; Lin, Jianping et al. (2014) AAV8 induces tolerance in murine muscle as a result of poor APC transduction, T cell exhaustion, and minimal MHCI upregulation on target cells. Mol Ther 22:28-41
Nagai, Y; Limberis, M P; Zhang, H (2014) Modulation of Treg function improves adenovirus vector-mediated gene expression in the airway. Gene Ther 21:219-24
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; Lipschutz, Joshua H (2014) Cilia and polycystic kidney disease, kith and kin. Birth Defects Res C Embryo Today 102:174-85
Myint, Melissa; Limberis, Maria P; Bell, Peter et al. (2014) In vivo evaluation of adeno-associated virus gene transfer in airways of mice with acute or chronic respiratory infection. Hum Gene Ther 25:966-76
Bryant, Laura M; Christopher, Devin M; Giles, April R et al. (2013) Lessons learned from the clinical development and market authorization of Glybera. Hum Gene Ther Clin Dev 24:55-64

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