In this application protocol, we describe a rapid protocol for the dissociation of neonatal mouse or rat hearts followed by the efficient immunomagnetic enrichment of cardiomyocytes, cardiac fibroblasts, and endothelial cells.
Dissociate neonatal heart tissue from mouse or rat with the Neonatal Heart Dissociation Kit. This kit enables enzymatic tissue dissociation followed by semi-automated mechanical tissue dissociation on the gentleMACS™ Dissociator. Follow the protocol of the kit data sheet.
Download data sheet
Use the Neonatal Cardiomyocyte Isolation Kit to enrich cardiomyocytes by depleting non-target cells. The kit contains a cocktail of monoclonal antibodies conjugated with MACS® MicroBeads that magnetically label non-target cells, which can then be depleted by retaining them within a MACS Column in the magnetic field of a MACS Separator, while the unlabeled cardiomyocytes pass through the column. Follow the protocol of the kit data sheet.
Download the data sheet
Use the Neonatal Cardiac Fibroblast Isolation Kit to enrich fibroblast cells in a two-step process that first labels and depletes non-fibroblast cells, and then selects fibroblast cells for elution. Follow the protocol of the kit data sheet.
Download the data sheet
Highly enriched endothelial cell populations are a prerequisite to facilitate vascularization in tissue-engineered myocardium. Isolate cardiac endothelial cells from dissociated neonatal heart tissue with the Neonatal Cardiac Endothelial Cell Isolation Kit. The kit enables a two-step process that first labels and depletes non-endothelial cells, and then selects and retains endothelial cells in the column for subsequent elution. Follow the protocol of the kit data sheet.
Download the data sheet
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autoMACS, CliniMACS, the CliniMACS logo, CliniMACS Prodigy, CryoMACS, CytoMix, CytoStim, DendriMACS, ExiTron, ExpAct, FeraSpin, FeraTrack, GadoSpin, gentleMACS, LIFE 18, LIFE 21, MACS, the MACS logo, MACSductin, MACSelect, MACSfectin, MACSflex, MACSiBead, MACSiMAG, MACSmix, MACSprep, MACSQuant, MACSQuantify, MACSxpress, MidiMACS, MiniMACS, miRXplore, MultiMACS, NeuroBrew, NiraWave, OctoMACS, PepTivator, pMACS, PolySon, PrepProtect, QuadroMACS, REAfinity, REAlease, Rheo, StemMACS, StraightFrom, SuperAmp, SuperMACS, TexMACS, TheraSorb, thermoMACS, TransAct, Tyto, the Tyto logo, VarioMACS, Vio, Viobility, VioBlue, VioBright, VioGreen, Viscover, and µMACS are registered trademarks or trademarks of Miltenyi Biotec B.V. & Co. KG and/or its affiliates in various countries worldwide. Ficoll-Paque is a trademark of GE Healthcare companies. Vectofusin-1 is a registered trademark of Genethon. All other trademarks mentioned in this document are the property of their respective owners and are used for identification purposes only. Copyright © 2019 Miltenyi Biotec B.V. & Co. KG and/or its affiliates. All rights reserved.
▲ Note: Addition of DNase (230 U/mL) to the buffer is optional, but may improve plating of endothelial cells after cell separation.
▲ Note: BSA can be replaced by other proteins such as mouse serum albumin, mouse serum, or fetal bovine serum (FBS). Buffers or media containing Ca2+ or Mg2+ are not recommended for use.
MACS Columns and MACS Separators: The recommended columns for optimal purity and recovery are listed in the table below. Depletion can also be performed using the autoMACS Pro Separator (mouse) or the MultiMACS™ Cell24 Separator (rat).
Column | Max. number of labeled cells | Max. number of total cells | Separator |
---|---|---|---|
Depletion | |||
Mouse | |||
MS | 5×106 | 1×107 | MiniMACS™, OctoMACS™, VarioMACS, SuperMACS II |
autoMACS® | 5×106 | 1×107 | autoMACS Pro |
Rat | |||
LD | 1×107 | 2×107 | MidiMACS™, QuadroMACS™, VarioMACS, SuperMACS II |
Multi-24 | 1×107 | 2×107 | MultiMACS Cell24 |
▲ Note: Column adapters are required to insert certain columns into the VarioMACS™ or SuperMACS™ II Separators. For details refer to the respective MACS® Separator data sheet.
Column | Max. number of labeled cells | Max. number of total cells | Separator |
---|---|---|---|
Depletion | |||
Mouse | |||
MS | 5×106 | 1×107 | MiniMACS, OctoMACS, VarioMACS, SuperMACS II |
autoMACS® | 5×106 | 1×107 | autoMACS Pro, autoMACS |
Rat | |||
LS | 1×107 | 2×107 | MidiMACS, QuadroMACS, VarioMACS, SuperMACS II |
autoMACS® | 1×107 | 2×107 | autoMACS Pro, autoMACS |
▲ Note: Column adapters are required to insert certain columns into the VarioMACS or SuperMACS II Separators. For details refer to the respective MACS Separator data sheet.
Column | Max. number of labeled cells | Max. number of total cells | Separator |
---|---|---|---|
Mouse | |||
Depletion | |||
LD | 1×108 | 5×108 | MidiMACS, QuadroMACS, VarioMACS, SuperMACS II |
Positive selection | |||
MS | 1×107 | 2×108 | MiniMACS, OctoMACS, VarioMACS, SuperMACS II |
Positive selection or depletion | |||
autoMACS® | 2×108 | 4×109 | autoMACS Pro, autoMACS |
Rat | |||
Depletion | |||
LD | 1×107 | 2×107 | MidiMACS, QuadroMACS, VarioMACS, SuperMACS II |
Positive selection | |||
MS | 1×107 | 2×107 | MiniMACS, OctoMACS, VarioMACS, SuperMACS II |
▲ Note: Column adapters are required to insert certain columns into the VarioMACS or SuperMACS II Separators. For details refer to the respective MACS Separator data sheet.
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