Fitc conjugated anti human cd90 thy1 antibody

Cells were seeded (2.0 × 10⁵ cells/well) on laminin (120 μg/mL) coated 12 well plates, after which differentiation was induced. Cells were harvested on the 0, 4, 9, and 14 days of differentiation and were suspended in 1% FBS in PBS. Cells were stained with the FITC-conjugated anti-human CD90 (Thy1) antibody (Cat.#. 328107; BioLegend Inc., San Diego, CA, USA) at 4 °C for 30 min in the dark. Cells were washed twice with 1% FSB in PBS and were analyzed using a FACS LSR II equipped with FACSDiva™ Software (Becton Dickinson Biosciences, San Jose, CA, USA).

hDPSCs were characterized according cell morphology, stemness and proliferative mRNA marker expression, and surface marker analysis. Cell morphology was captured by phase-contrast microscope. RT-qPCR was used to analysed mRNA marker expression regarding stemness property (REX1, NANOG, and OCT4) and proliferative marker (Ki67). Cells were then characterized by flow cytometry for MSC surface markers. Briefly, the cells were stained with FITC-conjugated anti-human CD105⁺ antibody (Bio Legend, California, USA), FITC-conjugated anti-human CD73⁺ (Ecto-5′-nucleotidase) antibody (Bio Legend, California, USA), FITC-conjugated anti-human CD90⁺ (Thy1) antibody (Bio Legend, California, USA), FITC-conjugated anti-human CD44⁺ antibody (Bio Legend, California, USA), and FITC-conjugated anti-human CD45⁻ antibody (Bio Legend, California, USA). FITC-conjugated Mouse IgG1κ isotype control (FC) antibody (Bio Legend, California, USA) was used as an isotype control for this assay. The assay was performed by using FACScallibur flow cytometer with CellQuest software (BD Bioscience, New Jersey, USA).