Microfabrication of low-cost customisable counting chambers for standardised estimation of sperm concentration

The creation of single and multilayered adult stem cells (ASCs) sheets is presented. The stem cell sheets preserve the cell-cell and cell-extracellular matrices and are developed by utilizing a thermally reversible methylcellulose (MC) coated tissue culture polystyrene (TCPS) dish. This technique is an improvement and a simplification of earlier noninvasive cell retrieval methods based on the use of a temperature-responsive poly(N-isopropylacrylamide) (PIPAAm) coated TCPS dishes. The optimal combination of MC-water-salt was determined to be 12-14% of MC (mol. wt. of 15,000) in water with 0.5× PBS (~150 mOsm). This solution exhibited a gel formation temperature of ~32 °C. The addition (evenly spread) of 1 ml of 3 mg/ml rat tail type-I (pH adjusted to 7.5) over the MC coated surface at 37 °C improves ASC adhesion and proliferation on the methylcellulose system. Upon confluence, a continuous monolayer ASC sheet was formed on the surface of the MC hydrogel system. When the grown cell sheet was removed from the incubator and exposed to room temperature (~30 °C), it spontaneously and gradually detached from the surface of the thermoresponsive hydrogel, creating an ASC sheet

Methylcellulose Based Thermally Reversible Hydrogels

The creation of single and multilayered adult stem cells (ASCs) sheets is presented. The stem cell sheets preserve the cell-cell and cell-extracellular matrices and are developed by utilizing a thermally reversible methylcellulose (MC) coated tissue culture polystyrene (TCPS) dish. This technique is an improvement and a simplification of earlier noninvasive cell retrieval methods based on the use of a temperature-responsive poly(N-isopropylacrylamide) (PIPAAm) coated TCPS dishes. The optimal combination of MC-water-salt was determined to be 12-14% of MC (mol. wt. of 15,000) in water with 0.5× PBS (~150 mOsm). This solution exhibited a gel formation temperature of ~32 °C. The addition (evenly spread) of 1 ml of 3 mg/ml rat tail type-I (pH adjusted to 7.5) over the MC coated surface at 37 °C improves ASC adhesion and proliferation on the methylcellulose system. Upon confluence, a continuous monolayer ASC sheet was formed on the surface of the MC hydrogel system. When the grown cell sheet was removed from the incubator and exposed to room temperature (~30 °C), it spontaneously and gradually detached from the surface of the thermoresponsive hydrogel, creating an ASC sheet