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CC161-1050UG ECMatrix™-511 Silk E8 Laminin Substrate

Xeno-free laminin-511 coating for feeder-free pluripotent stem cell cultures, 1050 µg (Silkworm derived)

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CC161-1050UG
1050 µg  
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      Overview

      Replacement Information
      Description
      Catalogue NumberCC161-1050UG
      DescriptionECMatrix™-511 Silk E8 Laminin Substrate
      Overview“The ECMatrix™ substrate has enabled a simpler workflow for human iPSC processing. We save time by eliminating the need to pre-coat our cultureware, while maintaining high-quality pluripotent stem cell cultures”. Research Scientist, Cell Therapy Bioprocessing Group.

      “Human ES cells grown on the ECMatrix™ substrate have high cell viability, proliferation and were able to differentiate after multiple passages. Additionally, the substrate saves us time by eliminating the need to pre-coat cultureware". Senior Stem Cell Scientist, Regenerative Medicine Process Development Group.

      Human pluripotent stem cells (ES and iPS cells) express α6β1 as the major integrin species and therefore can be maintained stably and expanded efficiently in feeder-free conditions on culture vessels coated with it’s binding partner laminin-511. However, laminin-511 is not suitable for large-scale production because of its large molecular weight and heterotrimeric nature. Professor Kiyotoshi Sekiguchi’s group (Matrixome, Inc.) have solved this problem by producing a recombinant E8 fragment of laminin-511 at large-scale while retaining the full integrin binding activity.1 The ECMatrix™-511 E8 Laminin Substrate can be used to culture pluripotent stem cells in feeder-free conditions with numerous added benefits over traditional methods including:

      Features and Benefits
      • Animal-free, xeno-free format: Consistent from lot-to-lot with no prescreening required
      • No plate precoating required: Save time by simply adding to media while passaging cells
      • Supports single cell passaging w/out ROCKi: Great for CRISPR editing or clonal isolation
      • Higher adhesion and growth rates: Get to your experiments faster
      • Easy to handle: No chilling of cell culture consumables required

      References
      1. Miyazaki T, et al. Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells. Nature Commun. 2012;3:1236.
      2. Nakagawa M, et al. A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells. Sci Rep. 2014 Jan 8;4:3594.
      3. Takashima Y, et al. Resetting transcription factor control circuitry toward ground-state pluripotency in human. Cell. 2014 Sep 11;158(6):1254-1269.
      4. Miyazaki T, et al. Efficient Adhesion Culture of Human Pluripotent Stem Cells Using Laminin Fragments in an Uncoated Manner. Sci Rep. 2017 Jan 30;7:41165.
      Alternate Names
      • iMatrix-511 SILK
      Background InformationProtocol

      Depending on application, either a precoating or non-precoating method can be used to culture pluripotent stem cells.

      Non-Precoating Method
      1. Detach cells into small clumps or single cells using Accutase.
      2. Add ECMatrix™-511 to fresh media at a final concentration of 0.25 ug/cm2 (for example: for one well of a 6-well plate add 5 uL of the 0.5 mg/mL stock solution).
      3. Add cells to the ECMatrix-511™/Media and plate the cells at desired density.

      Precoating Method
      1. Dilute the 0.5 mg/mL stock solution with sterile PBS to achieve a 2.5 ug/mL working solution.
      2. Coat dishes with ECMatrix™-511 at 0.25 ug/cm2 (for example, for one well of a 6-well plate add 1 mL of the 2.5 ug/mL working solution).
      3. Incubate for 1 hour at 37°C, 3 hours at room temperate or overnight at 4°C.
      4. Before use, remove remaining fluid from the coated surface (do not rinse).
      5. Detach cells into small clumps using Accutase.
      6. Plate the cells at desired density.

      Note: Do not allow the plates to dry, briefly spin down all liquids in the tube before use, avoid repeated freeze-thaw cycles.
      References
      Product Information
      Components
      • 1) 6 X 175 µg ECMatrix™-511 E8 Laminin Substrate (0.5 mg/mL in PBS). Expressed in transgenic silkworm cocoon.
      Applications
      ApplicationXeno-free laminin-511 coating for feeder-free pluripotent stem cell cultures, 1050 µg (Silkworm derived)
      Key Applications
      • Stem Cell Culture
      Biological Information
      Media FormLiquid
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assurance• Purity (SDS-Page): > 95%
      • Endotoxin Test: ≤ 750 EU/mg
      • Mycoplasma Test: Negative
      • Sterility Test: Negative
      • Integrin Binding Assay (kDa): ≤ 10 nM
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsECMatrix™-511 E8 Laminin Substrates should be stored at 2-8°C. Avoid multiple freeze-thaw cycles and protect from light.
      Packaging Information
      Material Size1050 µg
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalog Number GTIN
      CC161-1050UG 04061841030225