Description
General description
“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.
Application
Xeno-free laminin-511 coating for feeder-free pluripotent stem cell cultures, 350 μg (CHO-S derived)
Features and Benefits
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:
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
Packaging
350 μg (2 x 175 μg)
Preparation Note
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 μg/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 μg/mL working solution.
2. Coat dishes with ECMatrix-511 at 0.25 μg/cm² (for example, for one well of a 6-well plate add 1 mL of the 2.5 μg/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.
ECMatrix-511 E8 Laminin Substrates should be stored at 2-8°C. Avoid multiple freeze-thaw cycles and protect from light.
Analysis Note
Purity (SDS-Page): > 95%
Endotoxin Test: = 750 EU/mg
Mycoplasma Test: Negative
Sterility Test: Negative
Integrin Binding Assay (kDa) = 10 nM
Other Notes
2 X 175 μg ECMatrix-511 E8 Laminin Substrate (0.5 mg/mL in PBS). Expressed in CHO-S cells.
Disclaimer
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.
