Generation of a human embryonic stem cell line stably expressing high levels of the fluorescent protein mCherry

Figure 2 Utility of the MEL2-mCherry line in various analyses of human embryonic stem cell behavior. Robust mCherry expression of the MEL2-mCherry line allows for analysis of cellular behavior such as (A) mobility on various substrates or (B) mode of colony formation after mixing with non-fluorescent parental MEL2 human embryonic stem (hES) cells or (C, D) following encapsulation into transparent microcarriers; E-M: Utility of the MEL2-mCherry line for the analysis of cellular adhesion to various substrates. Uniformity and robustness of mCherry expression, in combination with its uniform distribution throughout the cell, allows the use of fluorescence level distribution to quantify cell adhesion (E-G), to identify dead, unattached cells (yellow in H-J), and accurately determine the area of substrate coverage by live hES cells (K-M). From left to right, three different ECM molecules were tested. In this substrate set relative areas of surface coverage were 4.4%, 41.7% and 76.2%.