Pluripotent Stem Cell Cardiac Differentiation

The following matrices were assessed for the ability to support pluripotent growth and subsequent cardiac differentiation: 9 µg/cm² growth-factor reduced Matrigel (1:200, Corning) in DMEM/F12; 625 ng/cm² vitronectin peptide (Synthemax II-SC, 1:320, Corning) in ultrapure water (1:50 also tested); 1 µg/cm² full length recombinant human vitronectin (1:50, Primorigen) in D-PBS with CaCl₂ and MgCl₂; 2.5 µg/cm² laminin-521 (1:80, Biolamina) in DPBS; 2 µg/cm² truncated recombinant human laminin-511 iMatrix-511 (1:50, Iwai North America, Foster City, CA, USA) in DPBS; 1 µg/cm² rH E-cadherin (1:25, StemAdhere, Primorigen/Stemcell Technologies); and 10 µg/cm² fibronectin (1:20, EMD Millipore) in D-PBS. All were used at 2 mL per well of a 6-well (9.6 cm²). Matrices were assessed on both 6-well polystyrene tissue culture plates and untreated plates (both from Greiner). Also tested were Synthemax-T 6-well plates, and fibronectin mimetic plates (both from Corning) and 10 µg/cm² Pronectin (Sigma-Aldrich).

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Burridge P.W., Matsa E., Shukla P., Lin Z.C., Churko J.M., Ebert A.D., Lan F., Diecke S., Huber B., Mordwinkin N.M., Plews J.R., Abilez O.J., Cui B., Gold J.D, & Wu J.C. (2014). Chemically Defined and Small Molecule-Based Generation of Human Cardiomyocytes. Nature methods, 11(8), 855-860.

Publication 2014

Cadherin Cardiac Fibronectin Growth factor Human Human laminin 511 Laminin Matrigel Mgcl2 Peptide Polystyrene Stemcell Tissue Vitronectin

Corresponding Organization : Stanford University

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Variable analysis

independent variables

9 µg/cm^2 growth-factor reduced Matrigel (1:200, Corning) in DMEM/F12
625 ng/cm^2 vitronectin peptide (Synthemax II-SC, 1:320, Corning) in ultrapure water (1:50 also tested)
1 µg/cm^2 full length recombinant human vitronectin (1:50, Primorigen) in D-PBS with CaCl2 and MgCl2
2.5 µg/cm^2 laminin-521 (1:80, Biolamina) in DPBS
2 µg/cm^2 truncated recombinant human laminin-511 iMatrix-511 (1:50, Iwai North America, Foster City, CA, USA) in DPBS
1 µg/cm^2 rH E-cadherin (1:25, StemAdhere, Primorigen/Stemcell Technologies)
10 µg/cm^2 fibronectin (1:20, EMD Millipore) in D-PBS
Synthemax-T 6-well plates
Fibronectin mimetic plates (both from Corning)
10 µg/cm^2 Pronectin (Sigma-Aldrich)

dependent variables

Ability to support pluripotent growth
Ability to support subsequent cardiac differentiation

control variables

All matrices were used at 2 mL per well of a 6-well (9.6 cm^2)
Matrices were assessed on both 6-well polystyrene tissue culture plates and untreated plates (both from Greiner)

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