A cell culture module capable of cooling stimulus to collect cells efficiently on a metal culture substrate was developed. We evaluated the cell collection ratio and morphology of the collected cells. Following a cooling stimulus (0◦ C) for 20 min, the number of collected cells was increased by 50% compared to that collected after trypsin treatment without pipetting from the metal culture substrate. Following the cooling stimulus, cells were observed by fluorescence microscopy and scanning electron microscopy; the cell filopodia were shrunken compared to non-cooling-stimulated cells. Furthermore, the combination of collagenase and cooling stimulation resulted in the collection of a comparable number of cells as that obtained using only trypsin. Thus, cell proliferation was improved compared to that following trypsin treatment. Therefore, this method can be applied for culturing cells that are susceptible to trypsin damage.
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