Collecting Blood for ips Cell Reprogramming Europe
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1 MyCell Products Application Protocol Collecting Blood for ips Cell Reprogramming Europe Introduction: Blood Collection and Infectious Disease Testing All persons collecting and handling any blood samples should be appropriately trained regarding (A) the safe handling and administration of blood and blood components, (B) bloodborne pathogen safety and exposure controls (tasks and procedures to minimize human exposure to such pathogens), and (C) any and all other requirements under all applicable laws and regulations and guidance as to best safety practices. Note: For clarification on this protocol, and request the MyCell Products Webinar: Getting Started. A trained phlebotomist must collect blood samples using the Vacutainer tubes included in the MyCell Blood Donor Collection Kit. All blood samples will undergo infectious disease testing (IDT) after receipt by Cellular Dynamics International (CDI): Ship the fresh blood to CDI within 24 hours of collection. The blood must be determined free of HBV, HCV, and HIV. IDT Name Hepatitis B Surface Antigen Hepatitis C Antibody HIV Antigen and Antibody Relative Current Procedural Terminology (CPT) Code In the case of a positive IDT result, CDI will discard the donor s samples. Required Equipment and Consumables The following equipment and consumables are required for collecting blood. Item Equipment Tabletop Centrifuge with Proper Adaptors for Vacutainer Tubes and Capable of Maintaining 4 C 1 Vendor Multiple Vendors Collecting Blood for ips Cell Reprogramming - Europe 1
2 Item Consumables MyCell Blood Donor Collection Kit 2,3,4 Vendor Cellular Dynamics International (CDI) 1 CDI recommends using a USA E8 Fixed Speed Centrifuge (LW Scientific, Cat. No. E8C-U8AF-1503) or an equivalent centrifuge. 2 Do not discard any materials included in the kit as they may be used for sample identification and shipment. 3 Store the gel packs at 4 C until ready to use. Do not freeze the gel packs. 4 Store the Vacutainer tubes at room temperature and use before the expiration date printed on the tube. The kit includes Vacutainer Lithium Heparin (BD Biosciences, Cat. No ), CPT (BD Biosciences, Cat. No ), and SST (BD Biosciences, Cat. No ) tubes. Notes Workflow Blood samples are collected in the Vacutainer tubes: 1 Lithium Heparin tube for DNA extraction, 1 SST tube for IDT, and 4 CPT tubes for reprogramming. Fresh blood samples are shipped to CDI within 24 hours of collection. Methods: Collecting Blood Samples The MyCell Blood Donor Collection Kit includes the Vacutainer Lithium Heparin, SST, and CPT tubes for blood samples. The kit also includes the MyCell Donor Sample labels with CDI-assigned ID and project numbers for sample identification. The phlebotomist should collect 1 Lithium Heparin tube (4 ml/tube), 1 SST tube (4 ml/tube), and 4 CPT tubes (8 ml/tube) from each donor. Note: Store the Vacutainer tubes at room temperature until ready to perform blood collection. Use before the expiration date printed on the tube. 1. Label each Vacutainer tube with a CDI-provided MyCell Donor Sample label (Figure 1). Figure 1: Labeling Vacutainer Tubes Apply a CDI-provided MyCell Donor Sample label to each tube to ensure proper identification. 2 Cellular Dynamics International, Inc.
3 Notes 2. Collect the blood samples by venipuncture using standard blood drawing procedures. Immediately invert the Lithium Heparin tube 8-10 times to ensure complete mixing. Invert the CPT and SST tubes 5 times to ensure complete mixing. 3. Maintain the Lithium Heparin tube on ice during incubation and subsequent centrifugation of CPT and SST tubes. Incubate the CPT and SST tubes at room temperature for minutes. Note: See Appendix A for recommendations on properly handling and processing SST tubes. 4. Centrifuge the CPT and SST tubes at room temperature using the speed and duration, as specified in the table below, for the centrifuge s rotor type. Do not centrifuge the Lithium Heparin tube. Note: See the manufacturer s instructions for adaptors needed to centrifuge Vacutainer tubes. Note: Ensure the rotor is correctly balanced to achieve proper blood separation. Vacutainer Tube Speed (x g) Duration (min) Swinging Bucket Rotor Fixed Angle Rotor CPT Tube SST Tube Inspect the CPT and SST tubes to ensure proper blood separation was achieved (Figure 2). Note: If the blood did not separate into phases, ensure the rotor is balanced and centrifuge the tubes once more; however, be aware there will be a reduction in the number of PBMCs isolated. If the blood still does not separate, collect a fresh sample from the same donor. CPT Tube SST Tube Figure 2: Vacutainer CPT and SST Tubes after Blood Centrifugation The blood separates into phases upon centrifugation. 6. Maintain the Lithium Heparin, SST, and CPT tubes on ice until ready to package for shipment. Note: Samples will be maintained on gel packs during shipment. See the packaging and shipping instructions, below. Collecting Blood for ips Cell Reprogramming - Europe 3
4 Sample Shipment: Shipping Fresh Blood The MyCell Blood Donor Collection Kit includes the packaging materials. Ship fresh blood with 24 hours of collection by World Courier for overnight delivery to CDI (PRA Health Sciences). Please ensure arrival on or before the Friday of the same week. Do not ship samples on a Friday. 1. Equilibrate the 2 gel packs at 4 C overnight. Note: Maintain the gel packs at 4 C until ready to use. Do not freeze the gel packs. 2. Download and complete the CDI Shipping Manifest: Note: See Appendix B for an example of the form. 3. Package the samples (Figure 3): a. Place the insulated shipper in the outer shipping box. b. Add a layer of bubble wrap to the bottom of the insulated shipper. c. Wrap the Vacutainer tubes in absorbent paper and place in the small vial carton. d. Place the small vial carton containing the Vacutainer tubes in a sealable plastic bag and then in the insulated shipper. e. Add another layer of bubble wrap and place 2 cold gel packs on top of the bubble wrap before sealing the insulated shipper s lid. 4. Place a printout of the completed CDI Shipping Manifest in the outer shipping box and in the site s study binder. 5. Before sample shipment, the World Courier tracking number and the CDI Shipping Manifest to mycell@cellulardynamics.com and KirschbaumMarc@prahs.com. This notification allows CDI and PRA Sciences to track and prepare for delivery of the shipment. 6. Ship by World Courier for overnight delivery to the following address: PRA Health Sciences - Early Development Services Bioanalytical Laboratory Amerikaweg 18, 9407 TK, Assen, Netherlands Notes A B C D E Figure 3: Shipping Vacutainer Tubes Containing Fresh Blood Package the tubes for shipment as follows: (A) Place the insulated shipper with a layer of bubble wrap in the outer shipping box. (B) Wrap the tubes in absorbent paper and place in the small vial carton. (C) Place the small vial carton in a sealable plastic bag and then in the insulated shipper. (D) Add another layer of bubble wrap followed by 2 cold gel packs. (E) Seal the insulated shipper s lid. (Not shown) Place a printout of the completed CDI Shipping Manifest in the outer shipping box before sealing. 4 Cellular Dynamics International, Inc.
5 Notes Appendices Appendix A. Proper Handling of Vacutainer SST Tubes See the next page for the November 2005 issue of Tech Talk (courtesy of Becton, Dickinson and Company), which details how to handle and process Vacutainer SST tubes. Collecting Blood for ips Cell Reprogramming - Europe 5
6 6 Cellular Dynamics International, Inc. Notes
7 Notes Appendix B. Example CDI Shipping Manifest Collecting Blood for ips Cell Reprogramming - Europe 7
8 Notes Customer s Responsibilities CDI does not guarantee that you will obtain equivalent results from using icell or MyCell products as described herein or that such use will not infringe any intellectual property right(s) of any third party(ies). You are solely responsible for obtaining any licenses you may require for your specific research use(s) of the icell or MyCell products not expressly conveyed under CDI s terms and conditions of sale or other transfer of the icell or MyCell products to you. Conditions of Use For life science research use only. Trademarks icell is a registered trademark, and Cellular Dynamics and the logo are trademarks of Cellular Dynamics International, Inc. All other brands, product names, company names, trademarks, and service marks are the properties of their respective owners. Copyright Notice 2016 Cellular Dynamics International, Inc. All rights reserved. Revision History Version 1.0: December 2016 AP-MPBCEUR Cellular Dynamics International, Inc.
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