iMatrix-411 Recombinant Human Laminin-411 E8 Fragment Protein

- Overview
  - iMatrix-411 is a recombinant human laminin-411 E8 fragment protein expressed in Chinese Hamster Ovary (CHO)-S cells. iMatrix-411 contains the integrin-binding site of the laminin-411 molecule. iMatrix-411 is a useful cell culture substrate for proliferation and differentiation of vascular endothelial cells and bile duct epithelial cells. iMatrix411 is also useful for the culture of other cells adhering to laminin-411.
    
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    More Details
    
    Recombinant Laminin Fragments
- Properties
  - Source
    
    CHO-S cells
    
    Type
    
    Recombinant Proteins
    
    Storage
    
    Store at 2°C to 15°C, protect from light.Shelf life is 2 years.
    
    Concentration
    
    0.5 mg/mL
    
    Molecular Weight
    
    150 kDa
    
    Reconstitution
    
    Recombinant human laminin-411 E8 fragment protein in PBS(-)
    
    Activity
    
    The dissociation constant (Kd) for the binding with integrin α6β1 is 10 nM or less.
    
    * For research use only. Not intended for human use.
- Applications
  - Application
    
    Cell culture
    
    Application Description
    
    Efficient induction of endothelial progenitor cells from iPS cells
    
    By the following method, iMatrix-411 can be coated onto a culture vessel. The optimum coating density may differ by cell-type, cell-line, medium selected, or purpose. Insufficient coating density may result in the detachment of cells and varied cell conditions while the excessive coating density may lead to difficulty in detaching cells for passage.
    Determine the optimal coating density. 0.5 μg/cm2 is a standard but test between 0.1 and 1.5 μg/cm2.
    1) Dilute iMatrix-411 with PBS(-). Use the diluted iMatrix-411 immediately. To coat with 0.5 μg/cm2 onto a 6-well plate with 9.6 cm2/well, dilute 9.6 μL of iMatrix-411 with 2 mL of PBS(-) per well.
    2) Place the diluted iMatrix-411 into a culture vessel and incubate either at 37°C for 1 h, or at room temperature for 3 h, or at 4°C overnight.
    3) Aspirate the coating solution. Then, immediately seed your cells. Do not allow the coated surface to dry.
    *If you face difficulties in detaching cells for passage, re-adjust the conditions (e.g., reduce the coating density).
- Reference
  - 1.Ohta R. et al., Laminin-guided highly efficient endothelial commitment from human pluripotent stem cells. Scientific Reports 6:35680 / DOI: 10.1038/srep35680 (2016).
    2.Lammert E.,Cleaver O.,Melton D., Mechanisms of Development (Elsevier)120 (1) 59-64 (2003).
    3.Takayama K. et al., Laminin 411 and 511 promote the cholangiocyte differentiation of human induced pluripotent stem cells. Biochemical and Biophysical Research Commun. 474 (1): 91-96 (2016).
    4.Nishimura K. et al., Estradial facilitates functional integration of iPSC-derived dopaminergic neurons into striatal neuronal circuits via activation of integrin a5b1. Stem Cell Reports 6 (4): 511-524 (2016).
    5.Matsuno K. et al., Redefining definitive endoderm subtypes by robust induction of human induced pluripotent stemcells. Differentiation 2016.04.002.
    6.Hayashi R. et al., Co-ordinated ocular development ofrom human iPS cells and recovery of corneal function. Nature 531, 368-80 (2016),
    7.Sasaki K. et al., Robust in vitro induction of human germ celll fate from pluripotent stem cells. Cell Stem Cell 17 (2):178-194 (2015).
    8.Okumura N. et al., Laminin-511 and -521 enable efficient in vitro expansion of human corneal endothelial cells. Invest Ophthalmal Vis Sci. 56 (5), 2933-42 (2015).
    9.Nakagawa M. et al., A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells. Scientific Reports 4: 3594 (2014).
    10.Miyazaki T. et al. Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotentn stem cells. Nature Communications 3: 1236 (2012).
    11.Taniguchi Y. et al., The C-terminal region of laminin β-chains modulates the integrin-binding affinities of laminins: J. Biol. Chem. 284 (12): 7820-31 (2009).
    12.Ido H. et al., The requirement of the glutamic acid residue at the third position from the carboxyl termini of the laminin gamma-chains in integrin-binding by lamiinins. J. Biol. Chem. 282 (15): 11144-54 (2017).