ExCellerate™ iPSC Expansion Medium Animal-Free, GMP
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biotechne RDsystems ExCellerate™ iPSC Expansion Medium Animal-Free, GMP Catalog Number: CCM036-GMP Volume: 500 mL PRODUCT DESCRIPTION ExCellerate iPSC Expansion Medium, Animal Free, GMP is a complete, specifically formulated medium for in vitro feeder-free expansion of undifferentiated human pluripotent stem cells, including embryonic and induced pluripotent stem cells. This media was produced in a GMP animal free facility with no materials of animal or human origin used in the production of this medium or its components. STABILITY & STORAGE Upon receipt, store the medium at < -20 °C until the expiration date on the Certificate of Analysis. Thaw the medium overnight at 2-8 °C. Never thaw the medium at 37 °C. Upon thawing, the medium may be stored at 2-8 °C for two weeks. PRECAUTIONS When handling bio-hazardous materials, safe laboratory procedures should always be followed, and PPE should be worn. Acute and chronic effects of over-exposure to this medium are unknown. LIMITATIONS • For pre-clinical or ex vivo clinical use. • This reagent should not be used beyond the expiration date indicated on the Certificate of Analysis. • Results may vary due to variations among pluripotent stem cell populations derived from different donors. OTHER MATERIALS REQUIRED • Cell Adhesion Substrate such as Cultrex™ UltiMatrix (Catalog # BME001), ReadyBME (Catalog# 3434-050-RTU), or Vitronectin • Pipettes and pipette tips • 6 cm culture dishes • 15 mL conical tubes • 37 °C, 5% CO2 incubator • Inverted microscope • Cell Counter • Centrifuge • Cell harvesting solution (e.g. EDTA based such as Versene™ or Accutase®) • D-PBS • ROCK Inhibitor (Tocris™ Y-27632, Catalog # TB1254-GMP) Alternatively; CEPTCocktail available from Tocris, (Catalog # 7163), Emricasan (Catalog # 7310), Polyamine Supplementx1000 (lyophilized) (Catalog # 7739), and Trans-ISRIB (Catalog # 5284) Global bio-techne.com info@bio-techne.com TEL +1 612 379 2956 USA TEL 800 343 7475 Canada TEL 855 668 8722 Europe | Middle East | Africa TEL +44 (0)1235 529449 China TEL +86 (21) 52380373 Trademarks and registered trademarks are the property of their res

PROCEDURE FOR THE EX VIVO CULTURE OF HUMAN INDUCED PLURIPOTENT STEM CELLS The protocols below describe the expansion of human pluripotent stem cells using ExCellerate™ iPSC Expansion Medium, Animal Free, GMP in combination with either Cultrex™ Basement Membrane Extract (BME) or Recombinant Human Vitronectin. This protocol is optimized for 6 cm culture dish using the ROCK Inhibitor Y-27632 (Tocris™, Catalog #1254) but can be adjusted to use other culture vessels and ROCK inhibitors. 1. Prepare a 6 cm dish coated with either Cultrex Ultimatrix, ReadyBME, or Vitronectin. 2. Warm only the...

CLUMP PASSAGING OF iPSCs For regular subculturing, we recommend performing clump passaging of pluripotent stem cells. 1. Prepare coated 6 cm dishes. 2. Warm ExCellerate™ iPSC Expansion Medium and an EDTA solution (such as Versene™) at room temperature for 20-30 minutes. 3. Aspirate cell media, wash once with 1 mL of EDTA solution or D-PBS and then add 1 mL of EDTA solution and incubate at room temperature for approximately 5 minutes. 4. Remove the EDTA solution and add 1 mL of iPSC expansion media. Note: Most cells should still be lightly adherent to the dish and only lift off once medium...

END USER TERMS OF USE OF PRODUCT Production records and facilities are available for examination by appropriate personnel on site at R&D System in Minneapolis, Minnesota USA. R&D System sells GMP products for preclinical or clinical ex vivo use. They are not for in vivo use. Read the following End of Terms prior to using this product. The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and...