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  • Continuous harvest of stem cells via partial detachment from thermoresponsive nanobrush surfaces.

Continuous harvest of stem cells via partial detachment from thermoresponsive nanobrush surfaces.

Biomaterials (2015-11-01)
I-Chia Peng, Chin-Chen Yeh, Yi-Tung Lu, Saradaprasan Muduli, Qing-Dong Ling, Abdullah A Alarfaj, Murugan A Munusamy, S Suresh Kumar, Kadarkarai Murugan, Hsin-chung Lee, Yung Chang, Akon Higuchi
ABSTRACT

Stem cell culture is typically based on batch-type culture, which is laborious and expensive. Here, we propose a continuous harvest method for stem cells cultured on thermoresponsive nanobrush surfaces. In this method, stem cells are partially detached from the nanobrush surface by reducing the temperature of the culture medium below the critical solution temperature needed for thermoresponse. The detached stem cells are harvested by exchange into fresh culture medium. Following this, the remaining cells are continuously cultured by expansion in fresh culture medium at 37 °C. Thermoresponsive nanobrush surfaces were prepared by coating block copolymers containing polystyrene (for hydrophobic anchoring onto culture dishes) with three types of polymers: (a) polyacrylic acid with cell-binding oligopeptides, (b) thermoresponsive poly-N-isopropylacrylamide, and (c) hydrophilic poly(ethyleneglycol)methacrylate. The optimal coating durations and compositions for these copolymers to facilitate adequate attachment and detachment of human adipose-derived stem cells (hADSCs) and embryonic stem cells (hESCs) were determined. hADSCs and hESCs were continuously harvested for 5 and 3 cycles, respectively, via the partial detachment of cells from thermoresponsive nanobrush surfaces.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Collagenase from Clostridium histolyticum, for general use, Type I, ≥125 CDU/mg solid
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium - low glucose, With 1000 mg/L glucose and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Poly(ethylene glycol) methyl ether methacrylate, average Mn 500, contains 100 ppm MEHQ as inhibitor, 200 ppm BHT as inhibitor