Viastain aopi staining kit

Cell count, cell viability, and cell size were determined with the Cellometer Auto 2000 Cell Viability Counter using the ViaStain AOPI staining kit (both from Nexcelom Bioscience, Waltham, MA, USA). The exponential growth phase of the cells was estimated from a line created by plotting the natural logarithm of the cell number against the culture time. The slope of this linear curve is determined by linear regression analysis (GraphPad Prism, Version 5.01, GraphPad Software Inc.), which represents the maximal growth rate.

The growth kinetic of five cell populations from different cords (two male donors, three female donors) in 2D was investigated in a 12-well plate (CytoOne, USA Scientific, Item number CC7682-7512) to determine the growth parameter and as a comparison to the cell growth in the dynamic system. The cells (passage 3) were seeded at a density of 8000 cells·cm⁻² and a working volume of 1 mL DMEM LG medium. The cells were cultured for 7 days and harvested. To harvest, the medium of three wells was aspirated, washed once with 1 mL Dulbecco's phosphate buffered saline (DPBS), and lifted from the substrate with 0.05% trypsin-EDTA (Gibco, Cat. number 25200-056). The cell number, cell viability, and cell size were determined with the Cellometer Auto 2000 Cell Viability Counter using ViaStain AOPI staining kit (both from Nexcelom Bioscience, Waltham, MA).