Aggregate survival tests <i>in vitro</i>.

<p><b>A)</b> Subsets of microwell arrays with 2D monolayer of cell culture (2D) and aggregates of three sizes (S, M, and L). Hydrogen peroxide and anoxia/reoxygenation treatments were employed to induce cell death. EthD (red) and DAPI (blue) staining were performed for the determination of cell death. <b>B)</b> Quantification of dead CSP cells in 2D single layer culture and aggregates with variable diameters subjected to 200 µM-hydrogen peroxide treatment using EthD/DAPI fluorescent intensity ratio. Data were normalized to the vehicle groups of 2D monolayer culture and aggregates in three sizes. <b>C)</b> Quantification of dead CSP cells in 2D single layer culture and aggregates with variable diameters subjected to anoxia/reoxygenation using EthD/DAPI fluorescent intensity ratio. Data were normalized to the vehicle groups of 2D monolayer culture and aggregates in three sizes.</p

Cell patterning and aggregate formation inside microwells.

<p><b>A)</b> Cell patterning. Cells were localized inside the microwells. <b>B)</b> After cell seeding, the cells in the microwell array were cultured in a petri dish and aggregates formed within 24 h. <b>C)</b> Once the aggregate formation is complete inside the microwells, they can be stained. <b>D)</b> Aggregates can be imaged inside microwells. <b>E)</b> Aggregates can be easily released from the microwells by gentle flushing with media for other applications.</p

Aggregate integrity and survival in fluidic manipulations.

<p><b>A)</b> Aggregates formed in microwells can be easily flushed out from the microwell and centrifuged while remaining intact. <b>B)</b> Aggregate can be easily passed through a 30G needle without loosing integrity. <b>C)</b> A representative DAPI/EthD fluorescent image of aggregates before injection. <b>D)</b> A representative DAPI/EthD fluorescent image of aggregates after injection. <b>E)</b> Quantification of dead CSP cells in aggregates passing a 30G needle using EthD/DAPI fluorescent intensity ratio. (All bars represent 100 µm).</p

CSP cell survival <i>in vivo</i> following cardiac injury.

<p><b>A)</b> Protocol to measure the <i>in vivo</i> survival of CSP aggregates and suspensions. <b>B)</b> Representative serial bioluminescence images (BLI) of mice injected with CSP cell aggregates and CSP single cell suspensions. <b>C)</b> Percentage of CSP cell survival measured with BLI.</p