iPSCs and iPSC-derived cells were harvested with Accutase (Gibco, Thermo Fisher Scientific) and resuspended in ice-cold ocular differentiation medium. Cells were filtered through a 40 μm pore size cell strainer (Corning) and stained with Zombie UV Fixable Viability Kit (Biolegend) for 10 min on ice. After washing, samples were treated with antibodies on ice for 30 min and washed before acquisition on BD Fortessa. Results were analyzed using the FlowJo software (TreeStar, San Carlos, CA). Antibodies used for flow cytometry are depicted in Supplementary Table 3 .
40 μm pore size cell strainer
Manufactured by Corning
Sourced in United States
The 40 μm pore size cell strainer is a versatile lab equipment used for the filtration and separation of cells and cell aggregates. It features a pore size of 40 micrometers, which allows for the efficient isolation and purification of various cell types from complex samples.
Automatically generated - may contain errors
2 protocols using 40 μm pore size cell strainer
1
Isolation and Characterization of iPSC-Derived Cells
2
Sperm Purification and RNA Extraction
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To reduce the viscosity of the collected samples, 5 ml PBS (with 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4 and 1.8 mM KH2PO4, at pH 7.4) was added to the collected semen samples and the tubes were inverted to gently mix the contents before filtering with a 40 μM pore size cell strainer (Corning, Harrodsburg, KY, USA) on ice. After centrifugation at 800×g for 10 min at 4°C, the sperm were resuspended in 2 ml PBS, and 10 ml of somatic cell lysis buffer (0.1% sodium dodecyl sulphate and 0.5% Triton X-100 in diethylpyrocarbonate water) was added. The samples were incubated for 20 min on ice then centrifuged for 10 min at 4°C at 800×g, and the precipitated sperm were washed twice with 3 ml PBS to remove the somatic cell debris and then centrifuged again for 10 min at 4°C at 800×g. The purified sperm were lysed with 1 ml TRIzol (Invitrogen, Waltham, MA, USA) for RNA extraction.
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