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Human Platelet Lysate Faqs UPDATED FEB20

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Human Platelet Lysate Faqs UPDATED FEB20 LSP-DOC-3339 (ID 11862) Human Platelet Lysate FAQs.docx Frequently Asked Questions (FAQs): Human Platelet Lysate What is hPL? Human Platelet Lysate (hPL) is a cell culture growth supplement obtained from human platelets. hPL contains abundant human growth factors including PDGF, bFGF, EGF, TGF-beta1, VEGF, other cytokines and proteins which yield superior results when culturing many types of human cells, particularly human MSC, regardless of their tissue source. Serum (traditionally FBS or Human) is the supernatant of clotted blood, hPL is manufactured from lysed platelets. How is it collected? hPL is derived from human donor platelets collected from healthy consented volunteer donors at certified blood banks. All individual donors and complete batches are fully tested for pathogens according to 21CFR610. What are the benefits to using hPL instead of FBS? Performance: The future of cell therapy is to be xenogeneic-free and therefore human platelet lysate offers xenogeneic free/serum substitutes for replacing FBS and improving cell culture performance. Cost: FBS is commonly used at concentration between 10-20% in cell culture. However due to the high performance effects of human platelet lysate it is typically used at lower concentrations; 2.5%, 5% or 10%. Batch-to-batch variation: Batch-to-batch variability of hPL is significantly lower than that of FBS because it is derived from standardized human blood products. Since hPL is obtained from multiple donors units that are subsequently pooled in large batch sizes, it enables the manufacture of a consistent product with minimal lot-to-lot variation. What concentration of hPL should I use? Human Platelet Lysate (hPL) can be used at a variety of concentrations, from 10% in basal media to as little as 2.5%. The table below indicates the concentrations of hPL used for a variety of different cell types and applications. Which cell types has hPL been shown to work with? Concentration Species Cell Type Cell Name Description Used (%) Bovine Primary Cell CEC Corneal Endothelial Cells 2.5 - 10% Hamster Cell Line CHO Chinese Hamster Ovary epithelial Unknown Human Cell Line HEK-293 Human Embryonic Kidney 293 cells 1 - 10% Human Cell Line HeLa HeLa Cervical cancer cells 10% Human Cell Line HFFF2 HFFF2 Dermal fibroblasts 5 - 20% Human Cell Line HL-60 Human leukaemia cell lines 20% Human Cell Line HUVEC Human umbilical vein endothelial cells 5 - 20% Page 1 of 8 Life Science Production, a Division of Life Science Group Ltd. Tel: +44 (0) 1234 889180; Email: [email protected] Web: www.lifesciencegroup.co.uk LSP-DOC-3339 (ID 11862) Human Platelet Lysate FAQs.docx Human Cell Line JURKAT Human leukaemia cell lines 10% Human Cell Line K562 Myelogenous leukaemia 10% Human Cell Line KG-1 Human leukaemia cell lines 20% Human Cell Line MCF-7 Human continuous tumour 10% Human Cell Line NCTC 2544 Dermal keratinocytes 5% Human Cell Line NCTC 2544 Dermal keratinocytes 5 - 20% Human Cell Line A-549 Human continuous tumour 5% Human Cell Line Caco-2 Human continuous tumour 5% Human Cell Line HaCaT Keratinocyte cell line from adult human skin 5% Human Cell Line HEK-293 Human Embryonic Kidney 293 cells Unknown Human Cell Line HGF-1 Human Gingiva Fibroblasts 5% Human Cell Line MCF-7 Human continuous tumour 10% Human Cell Line U-251 MG Human continuous tumour 5% Mouse Cell Line C2C12 Myoblastoma Cells 1 - 20% MMT Mouse Cell Line Mouse mammary tumour Unknown 060562 Mouse Cell Line Neuro-2a Neuroblastoma cell line Unknown Mouse Cell Line NIH/3T3 Mouse Embryonic Fibroblasts 1 - 10% Mouse Cell Line RAG Mouse adenocarcinoma cell line Unknown Rabbit Cell Line SIRC Statens Seruminstitut Rabbit Cornea 1 - 10% Rat Primary Cell MSC Mesenchymal Stem Cells 2.5 - 5% Rat Cell Line R2C Testis from rat Unknown The above list represents cell lines and conditions that have been stated in published literature or from customer testimonials. We would recommend testing your cells with a titration range to determine the ideal conditions for your particular cells and application. What does it mean when hPL is labelled as Research Grade, GMP or GMP Clinical Grade? Research Grade material is manufactured to meet the needs of basic research and development. This material supports the in vitro propagation and maintenance of various human cell types. In addition, it provides a cost effective alternative to using serum-free media. Material is supplied in validated PETG NALEGENE bottles (25 mL, 100 mL and 500 mL). GMP Grade material is manufactured under GMP-controlled conditions for use in GMP compliant cell culture SOPs. This material is provided in cryo-bags (100 mL) or in validated PETG Nalgene bottles (100 mL and 500 mL). GMP Clinical Grade material is manufactured and gamma irradiated under GMP conditions to comply with the high safety guidelines for clinical applications. Due to pathogen-reduction, GMP Clinical Grade hPL provides a high safety level for expansion of different human cell types (particularly adult stem cells, e.g. MSCs) for clinical and therapeutic applications. Page 2 of 8 Life Science Production, a Division of Life Science Group Ltd. Tel: +44 (0) 1234 889180; Email: [email protected] Web: www.lifesciencegroup.co.uk LSP-DOC-3339 (ID 11862) Human Platelet Lysate FAQs.docx Which hPL product should I choose? Product Key Benefits Pack sizes Fibrinogen Requires Animal- available -depleted heparin? component free Human Platelet Animal serum-free; 25 mL, 100 mL No Yes Yes* Lysate Solution most cost effective & 500 mL option Human Platelet Animal serum-free; 10 mL, 50 mL, Yes No No** Lysate -FD fibrinogen-depleted 100 mL & 500 mL Human Platelet Animal component- 10 mL, 50 mL, Yes No YesƗ Lysate-XF free; fibrinogen- 100mL & 500 depleted mL Human Platelet Animal serum-free; Customisable No Yes Yes* Lysate - Customised customisable; small scale process development *does not contain any heparins (these are added at the point of use). **contains residues of porcine- derived heparin. ᵻ contains residues of fully synthetic heparin. Which Heparin product should I use? Only the Human Platelet Lysate Standard and Customised products requires the addition of heparin. There are two types of heparin available: 1. Heparin - Standard animal-derived heparin (5,000 U/mL) - HSUP-001M. 1. Xeno-Free Heparin - Animal component-free heparin, fully synthetic and preservative-free (12.5 mg/mL) - HSUP-XF-001M. What does ‘Xeno-Free’ mean? Our ‘Xeno-Free’ products are non-xenogeneic and animal serum-free. i.e. they do not contain material from species other than human origin. Materials that are described as being ‘Xenogenic’ are derived from a different species and therefore are not genetically or immunologically compatible. This is very important for cell therapy and clinical applications. What is the difference between ‘animal-serum free’ and ‘animal-component free’? All of our hPL products are derived from human platelets and therefore are naturally free form other animal derived material. No animal serum is added during the manufacture of these products. For a product to be animal-component free, no products can be added to the product during the manufacturing process that are derived from another species. For an animal-component free option (i.e. Page 3 of 8 Life Science Production, a Division of Life Science Group Ltd. Tel: +44 (0) 1234 889180; Email: [email protected] Web: www.lifesciencegroup.co.uk LSP-DOC-3339 (ID 11862) Human Platelet Lysate FAQs.docx free from non-human species), either choose Human Platelet Lysate Standard with Xeno-free heparin, or Human Platelet Lysate Solution XF (which is manufactured using Xeno-free heparin, a fully synthetic heparin product). What does it mean when hPL product is described as being Fibrinogen-depleted? For both Human Platelet Lysate-XF (Xeno-Free) and Human Platelet Lysate-FD (Fibrinogen Depleted), a fibrinogen depletion step is performed during the manufacturing process in order to remove all fibrinogen. In addition, heparin is added during the manufacturing process. This means that you do not need to add heparin while preparing the cell culture medium. For Human Platelet Lysate FD, the heparin used in manufacturing is derived from porcine intestine (and therefore not xeno-free). For Human Platelet Lysate XF, the heparin used is a xeno-free version, and is fully synthetic, offering a completely xeno-free (animal-component free) end product. Why is heparin not required for the FD and XF products? Heparin is an anti-coagulant that prevents the fibrinogen in human platelet lysate from clotting in the cell culture medium during cell expansion. Both Human Platelet Lysate XF and Human Platelet Lysate FD are fibrinogen-depleted and so do not require the addition of heparin in the cell culture media. What cell types are particularly recommended? Human Mesenchymal Stromal Cells from Bone Marrow (hMSC-BM) Human Mesenchymal Stromal Cells from Umbilical Cord (hMSC-UC) Human Mesenchymal Stromal Cells from Adipose Tissue (hMSC-AT) Human Dermal Fibroblasts (HDF) In addition, see FAQ ‘Which cell types has hPL been shown to work in?’, for a more detailed table of other cell types that hPL has been shown to work with. Can I use hPL in diagnostic or therapeutic applications? No. hPL is for in vitro experimental and research use only (RUO) and the product is not intended for human or animal diagnostic or therapeutic uses. What tests are carried out on hPL? Each batch of hPL is tested to ensure the absence of bacteria, fungi, yeast, mycoplasma (M. fermentans, M. arginini, M. orale, M. hyorhinis, M. salivarium, M. hominis, M. pneumoniae, Acholeplasma laidlawii, M. synoviae and Ureaplasma species). Batches are also tested for the ability to support growth of specific cell lines. In addition, each batch is also tested for standard parameters. These include endotoxin, pH, osmolality and total protein.
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