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. 2015 Jan 12;(95):52049. doi: 10.3791/52049
Name of equipment/ reagent Company Catalogue no. Comments/description
Cell culture Collagenase D Roche 11088866001
Dispase II Sigma D4693-1G Must be filter sterilized before use
Trypsin/EDTA (Gibco) Invitrogen 15400-054
100 µm cell strainer BD Biosciences 352360
Collagen solution ( 3 mg/ml in 0.1% acetic acid) Sigma, Dorset, UK C8919
Minisart SRP15 syringe filter (0.2 µm) Sartorius 17573ACK Polytetrafluorethylene (PTFE) membrane
CD56 human Microbeads Miltenyi Biotech 130-050-401 Magnetic-activated Cell Sorting (MACS) Be aware of the limited shelf life of microbeads
Anti- fibroblast Microbeads, human Miltenyi Biotech 130-050-601
40 μm Pre-separation filters Miltenyi Biotech 130-041-407
Large Cell Collumns Miltenyi Biotech 130-042-202 These columns come with a flow resistor. Use of the flow resistor is not necessary to obtain the high myogenic purities described here.
LS columns Miltenyi Biotech 130-042-401
MiniMacs Seperator Miltenyi Biotech 130-042-102 This separator fits the large cell column but not the LS column.
MidiMACS Miltenyi Biotech 130-042-302
MACS multistand Miltenyi Biotech 130-042-303
BSA Sigma Must be filter sterilized before use
Oil Red O Sigma O0625 Lipid staining
Triethyl phosphate Sigma 538728
Whatman Paper Sigma Z241121-1PAK No. 42, Ashless. To prepare the filter fold the circular filter paper to make a semi circle, then fold the semi-circle in half again to form a cone shape. Fit the cone into a funnel for filtering.
Mounting
ProLong Gold Antifade Reagent Molecular Probes, Invitrogen P36930 This can be purchased with or without DAPI and does not quench initial fluorescence.
AxioVision Carl Zeiss Contact Zeiss Image aquisition software
Adobe Photoshop CS5 Extended Adobe (purchased from Pugh Computers) ADPH16982* Image analysis software