Anti cd29

Conjugated antibodies against NANOG (phycoerythrin (PE) conjugated goat polyclonal (IC1997P), 1:200 dilution), OCT-3/4 (PE conjugated goat polyclonal (IC1759P), 1:200 dilution), and Nestin (PE conjugated mouse monoclonal (IC1259P), 1:200 dilution) were purchased from R&D Systems Europe Ltd (Abingdon, UK). Fluorescein isothiocyanate (FITC)-conjugated anti-CD90 (mouse monoclonal, 1:100 dilution) was purchased from Dianova GmbH (Hamburg, Germany).
All other antibodies used for cytofluorimetric analyses (FACS) were directly conjugated with FITC, PE, or APC: anti-CD31, antiCD-146/Muc-18, anti- CD29, anti-CD105, and anti-CD133 monoclonal antibodies (mAbs) (all from Miltenyi Biotec, Cologne, Germany); mAb anti-EpCAM (Biolegend, San Diego, CA). Isotype-matched FITC-, PE-, or APC- conjugated control mouse G (IgG) were from Miltenyi.

Induced pluripotent stem cell-derived-CMs were analyzed using a FACScan instrument (BD, Franklin Lakes, NJ, USA). The dead cells were identified with propidium iodide (PI, BD). Cell debris and dead cells were gated out, and the remaining cells were quantified for expression of eGFP using CellQuest v2.0 software (BD). The MSC surface markers were analyzed using the following fluorescently labeled antibodies: anti-CD29 (phycoerythrin, PE), anti-CD44 (biotin, anti-biotin-PE-Vio770), anti-CD90.2 (VioBlue), anti CD105 (allophycocyanin, APC), anti-Sca-1 (fluorescein isothiocyanate), and anti-CD11b (APC-Vio770) and fluorophore-matched isotype controls (all Miltenyi Biotec, Bergisch Gladbach, Germany). Cells were analyzed using a MACSquant flow cytometer and MACSquantify software (version: 2.4, both Miltenyi Biotec) with a 3% threshold (isotype control vs. specific antibody).