Abstract
Cell therapy usually accompanies cell detachment as an essential process in cell culture and cell collection for transplantation. However, conventional methods based on enzymatic cell detachment can cause cellular damage including cell death and senescence during the routine cell detaching step due to an inappropriate handing. The aim of the current study is to apply the pH-responsive degradation property of poly (amino ester) to the surface of a cell culture dish to provide a simple and easy alternative method for cell detachment that can substitute the conventional enzyme treatment. In this study, poly (amino ester) was modified (cell detachable polymer, CDP) to show appropriate pH-responsive degradation under mild acidic conditions (0.05% (w/v) CDP, pH 6.0) to detach stem cells (human adipose tissue-derived stem cells (hADSCs)) perfectly within a short period (less than 10 min). Compared to conventional enzymatic cell detachment, hADSCs cultured on and detached from a CDP-coated cell culture dish showed similar cellular properties. We further performed in vivo experiments on a mouse hindlimb ischemia model (1.0 × 106 cells per limb). The in vivo results indicated that hADSCs retrieved from normal cell culture dishes and CDP-coated cell culture dishes showed analogous therapeutic angiogenesis. In conclusion, CDP could be applied to a pH-responsive cell detachment system as a simple and easy nonenzymatic method for stem cell culture and various cell therapies.
Highlights
Stem cell detachment from cell culture dishes is an essential step in the cell culture process and cell collection for stem cell therapies [1,2]
Despite the prominent therapeutic results of stem cell therapies, conventional cell detaching methods based on enzymatic cell dissociation still contains intrinsic disadvantages such as stem cell death and senescence caused by an inappropriate handing [3,4]
We showed that we were able to retrieve the Human adipose tissue-derived stem cells (hADSCs) from cell culture dishes in an enzyme-free manner through a mild pH condition by applying CDP to the cell culture system
Summary
Stem cell detachment from cell culture dishes is an essential step in the cell culture process and cell collection for stem cell therapies [1,2]. Despite the prominent therapeutic results of stem cell therapies, conventional cell detaching methods based on enzymatic cell dissociation still contains intrinsic disadvantages such as stem cell death and senescence caused by an inappropriate handing [3,4]. Various trials including light- [5], temperature- [6,7], and pH-responsive [8,9,10] strategies for detaching stem cells from the cell culture dish without using enzymes have been reported. Previous reports usually require complicated steps, tailored equipment, and long operation time for stem cell detachment. A simple and easy method that can be suggested as an alternative method for enzymatic cell dissociation is required
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