Clonetics Cells in Pancreatic Cancer Research
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1 Pharma&Biotech BioResearch Clonetics Cells in Pancreatic Cancer Research 1 April 2014 / Speaker: Andrew Winner 2 April 2014 / Speaker: Dr. Nazim El-Andaloussi Lonza Cologne GmbH, Cologne / 28 March 2013 Lonza
2 Outline Introduction to Pancreatic Cancer Differences Between Primary Cells and Cell Lines Research Breakthroughs and Challenges Benefits of Clonetics Pancreatic Islets Custom Products for Pancreatic Research 2
3 Outline Introduction to Pancreatic Cancer Differences Between Primary Cells and Cell Lines Research Breakthroughs and Challenges Benefits of Clonetics Pancreatic Islets Custom Products for Pancreatic Research 3
4 Pancreatic Cancer Research Market Fourth leading cause of cancer deaths 1 Median survival time < 6 months 1 Challenging early stage detection NCI supports development of new treatments 2 1. Howard, 1966; Parker, 1996; Greenlee et al., Elekta 4 Mar-14
5 Pancreatic Cancer Types Neuroendocrine tumors (NET) Endocrine cancer Tumorous islets Ductal adenocarcinoma Exocrine Cancer Acinar cell carcinoma Rare malignant tumor; poor prognosis 5
6 Primary Cell Types Endocrine Region Pancreatic islets Alpha, beta cells, delta, PP, Epsilon 1 to 2% of pancreas Exocrine Region Acinar, ductal, centroacinar, stem cells Acinar cells 85% of pancreas Ductal cells 10% of pancreas 6
7 Outline Introduction to Pancreatic Cancer Differences Between Primary Cells and Cell Lines Research Breakthroughs and Challenges Benefits of Clonetics Pancreatic Islets Custom Products for Pancreatic Research 7
8 [Normal] Primary Cells vs. Pancreatic Cell Lines Primary Cells Cell Lines Islet Substitutions Mutation Rate Tissue Representation Characterization Need Costs Sample Number 8 Mar-14
9 Concerns About Cell Lines NIH - authentication of cultured cell lines critical for grant 1 Some journals mandate characterization before publication 18-36% cell lines misidentified or cross-contaminated 2 Discrepancies exist for phenotype and genotype of pancreatic cell lines to warrant careful scrutiny and thorough application of appropriate controls 3 1. Notice Number: NOT-OD <grants.nih.gov/grants/guide/notice-files/not-od html> 2. Hughes P et al. (2007) 3. Pancreas May; 39(4): Mar-14
10 Outline Introduction to Pancreatic Cancer Differences Between Primary Cells and Cell Lines Research Breakthroughs and Challenges Benefits of Clonetics Pancreatic Islets Custom Products for Pancreatic Research 10
11 Recent Research Breakthroughs First Human Model of Cancer Progression PSC s from Pancreatic Cancer Cells 1 Discovery of cancer stem cells in pancreatic tumors helps understand pancreatic cancer development 1. ScienceDaily June 20, 2013 < 11 Mar-14
12 Current Research Challenges Inconsistent literature on characterization profile of pancreatic cell lines 1 Limited cell line options for human islets 2 Limited access to normal primary human islets for research use Limited access to early-stage donor-cells 3 Isolation difficulties of primary cell types from pancreas 4 1. Pancreas May; 39(4): Gordon C. Weir. Clin Invest. 2011;121(9): Sharon Reynolds.Stem Cells Reveal Secrets About Cancer. 4. M.J Smelt et. al. Experimental Diabetes Research Volume 2008 (2008), Article ID , 11 pages 12 Mar-14
13 Outline Introduction to Pancreatic Cancer Differences Between Primary Cells and Cell Lines Research Breakthroughs and Challenges Benefits of Clonetics Pancreatic Islets Custom Products for Pancreatic Research 13
14 Why Use Clonetics Human Pancreatic Islets? Human islet center that manufactures for clinical transplantation Fresh and frozen Non-tumorous islets + acinar tissue Identity, quantity, purity, & viability testing Easy acquisition & steady supply 14
15 Why Use Clonetics Human Pancreatic Islets? Special donor characteristics Wilson Wolf Flask Growth Media included Technical Support 15
16 Clonetics High-Quality Islets Preparations Table: Characteristics of islet preparations and outcome of dynamic insulin function assay NHP (n=2) Human (n=3) AP (n=3) JP (n=4) Characteristics of Islet Preparations Insulin Secretory Function (AUC, pg insulin/ ng DNA) Purity (% dithizone stain) OCR/DNA (nmol/min.mg DNA) Total insulin (pg/ng DNA) Glucose 1 st + 2 nd phase (16.7 mm) 83 ± ± ± ± ± 57 h 121 ± 12 nhp,ap,jp 265 ± 25 h,jp 188 ± 3.2 a,h,ap 5.9 ± 0.9 jp 7.8 ± 3.9 jp 9.2 ± 3.4 jp 1.5 ± 0.8 nhp,h,ap 335 ± 7 ap,jp 348 ± 85 ap,jp 121 ± 78 nhp,h,jp 11.4 ± 4.6 nhp,h,ap a OCR/DNA data were available for 3 of 4 cased in the JP group. Data are presented as arithmetic means values ± SD. Statistical significance (P < 0.05): nhp compared with NHP; h compared with human; ap compared with AP; jp comapred with JP 16
17 Clonetics High-Quality Islets Transduction Control dsaav2-egfp Mouse Islets Human Islets Human and mouse islets transduced dsaav2-egfp virus Transduced islets were able to reverse diabetes In Vivo 17
18 Clonetics Islets and Western Blots Rat Human LacZ LacZ / ckd4 Cyclin D1 / lacz Cdk4 / cyclin D1 Uninfected LacZ LacZ / ckd4 Cyclin D1 / lacz Cdk4 / cyclin D1 Cyclin D1 Cdk4 Actin Western Blot Rat and human islets were transduced with different recombinant adenoviruses and assessed using immunoblots 18
19 Clonetics Fresh Human Pancreatic Islets Cat. No. Description Quantity Fresh Human Pancreatic Islets, 100K >=100,000 IEQ Fresh Human Pancreatic Islets, 20K >=20,000 IEQ Fresh Human Pancreatic Islets, 10K >=10,000 IEQ Fresh Human Pancreatic Islets, 5K >=5,000 IEQ Fresh Human Pancreatic Islets, 2K >=2,000 IEQ Special donor characteristics subject to donor availability; turnaround times may vary 19
20 Clonetics Fresh Human Pancreatic Islets Standard QC Testing Part Number Test Minimum Specs Guarantee Included Islet Quantity (IEQ Count) Included Islet Sterility Gram Negative Stain on Day 0 Included Islet Viability [FDA/PI] (%) >= 70% Included Islet Purity >= 70% islet cells vs. other tissue (based on visual representation) COA also includes age, sex, race & BMI of the donor Islets test negative for Hepatitis B, Hepatitis C & HIV-1 20
21 Clonetics Fresh Human Pancreatic Islets Additional QC Testing Cat. No. Test Typical Results (F.I.O.) Estimated Testing Time (Results Post-Shipment) DNA Quantitation /- 6.9 mg DNA 1 day Glucose Stimulated Insulin Response 4.1 +/- 2.8 fold 4 days Oxygen Consumption Rate / pmol/min/μg DNA 1 day Beta-Cell Composition /- 9.4% 14 days 21
22 Outline Introduction to Pancreatic Cancer Differences Between Primary Cells and Cell Lines Research Breakthroughs and Challenges Benefits of Clonetics Pancreatic Islets Custom Products for Pancreatic Research 22
23 Cells on Demand Human Fresh or Frozen Normal Acinar Tissue Frozen Normal Islets Pig, Monkey Fresh or Frozen Normal Islets and Acinar Tissue For additional requests not listed above, please contact our Cells on Demand Service Inquiries: 23 Mar-14
24 Lonza Cancer Product Portfolio Clonetics and Poietics Cells and Media tested and guaranteed to perform Primary cells from pancreatic, bone, blood, lymph, lung, mammary and other tissue types Nucleofector Technology for transfection of primary cells and cancer cell lines Clonetics Conditionally Immortalized Cell Lines (CCICs) Biowhittaker Liquid and Powder Media for cancer research and therapeutic applications Nucleic Acid and Protein Electrophoresis Products 24 Mar-14
25 Summary Restricted access to normal primary human islets and limited cell line options for research use Lonza s Clonetics Human Pancreatic Islets Fresh or frozen Steady supply Guaranteed quality Flexibility Lonza s Cells on Demand Service Acinar tissue Pancreatic cells from animal species 25
26 How to Order or Pose Questions Order queries: U.S : (toll free) or order.us@lonza.com Europe: or order.europe@lonza.com Scientific support: U.S: or scientific.support@lonza.com Europe: or scientific.support.eu@lonza.com Web address: for more information 26 Mar-14
27 Interested to Learn More? Join our upcoming webinar: Efficient Transfection of Biologically Relevant Cells in Immunology Research Slot 1: Tuesday, 13 May PM EDT (New York) / 11 AM PDT (Los Angeles) Slot 2: Wednesday, 14 May AM CEST (Berlin) / 9 AM BST (London) / 5 PM JST (Tokyo) Register at: 27
28 Thank You for Your Kind Attention
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