Perfusion Systems for Cell Isolation (PSCI) from Heart, Liver and Other Organs

The Perfusion System for Cell Isolation (PSCI) is specially designed for harvesting individual cells from isolated organs such as mouse, rat or guinea pig heart, liver and other organs. Individual cells are released from the cellular structure of the tissue through perfusion with an enzyme solution and are then flushed out.

Cell Isolation from Liver and Other Organs

Cardiomyocyte Isolation

As noted in the Item listing, you may purchase the base unit alone and add additional required items (see below; please consult technical support) or a complete functional system with all necessary components.

Cell Isolation from Liver and Other Organs

Features & Benefits

• Allows cell isolation from mouse, rat and guinea pig organs by enzymatic disintegration
• Specifically engineered dual perfusion system for blood cell flush and enzymatic disintegration
• Dedicated extension for cardiomyocyte isolation

Applications

• Harvesting individual cells from mouse, rat or guinea pig isolated organs 

Base units include: 

 Plexiglass stand, double heat exchanger, switching valve, protease reservoir and holder for pressure transducer. System volume: <3 ml (73-3659) or <5 ml (73-3639)

Additional equipment required: 

Thermocirculator, peristaltic pump, reservoir, magnetic stirrer, cannulae, operating table and others depending on version selected, transducers, monitoring system setup using the PLUGSYS Amplifier System.

Adaptation for Ex Vivo Perfusion

The PSCI can be adapted for ex vivo perfusion by adding A jacketed moist chamber and appropriate cannula for the organ of interest.

Adaptation for In Situ Perfusion

 The PSCI can be adapted for in situ perfusion by adding an operating table and appropriate cannula for the organ of interest.

Adaptation for Cardiomyocyte Isolation

See details below.

Setup and Operation

The system has two separate perfusion circuits and a specialized stopcock which allows the organ to be easily switched between the two circuits. The first circuit is filled with conventional perfusion solution and is used in the initial phase of the isolation process to flush out the blood cells from the organ. For the second phase, the system is switched to the second circuit which is filled with an enzyme solution for disintegration.

In the standard PSCI system, perfusion is performed under constant flow conditions. A peristaltic pump (purchased separately) is used to adjust the flow to a rate appropriate for the organ of interest. The system itself can handle flow rates of up to 50 or 100 ml/min, depending on the specific configuration. A pressure transducer and amplifier can easily be added to a set up to monitor perfusion pressure. Furthermore, the system can be upgraded to constant pressure perfusion with the addition of the SCP controller.

The easy to use system is designed to be as compact and user friendly as possible. Components that come in contact with perfusion solutions are alcohol resistant so that the perfusion circuits can be filled with alcohol to clean and sterilize after use.

The main system includes a dual heat exchanger and a platform for the peristaltic pump, both of which are secured to a vertical stainless steel rod and can be moved to appropriate positions for the process. Additional components are not fixed to the base as the dual heat exchanger and pump platform are. Instead, they are positioned on the PSCI baseplate and are connected to the system by tubing. The protease reservoir and the holder for the perfusion pressure transducer are included in the main system.

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Cardiomyocyte Isolation

Specially designed adaptation for harvesting individual cardiomyocytes from mouse, rat or guinea pig hearts

In case of using the heart version, a jacketed heart chamber is mounted on a slide which is clamped to the vertical column. A slow gas flow into this chamber ensure a positive pressure inside the heart chamber at all times, thus preventing ingress of bacteria from the surroundings during operation.

Features & Benefits

• Dual perfusion system for blood cell flushing and enzymatic digestion
• Compatible with disinfection using ethanol
• Positive pressure gas flow in heart chamber and reservoir prevent bacterial contamination
• Low priming volume conserves collagenase and minimizes temperature loss through tubing (<3 ml for mouse system, <5 mL for rat system)
• Integrated platform for roller pump minimizes system footprint and conserves use bench space
• Multi-purpose system can be fitted for use with other organs (e.g. in situ or ex vivo hepatocyte isolation from rat or mouse liver)

Applications

• Cardiomyocyte isolation
• Monitoring perfusion pressure
• Monitoring coronary flow

Complete Functional Systems

Base units include: 

Plexiglass stand, double exchanger, switching valve, heart chamber, aortic cannula OD 1.0 mm (mouse) or 2.0 mm (rat), protease reservoir and holder for pressure transducer. 

Additional equipment required if purchased separately: 

Thermocirculator, analog roller pump and Tygon tubing, aortic cannulae (1.0 mm OD for mouse; 2.0, 2.5, or 3.0 mm OD for rat/guinea pig), protease reservoir, mini magnetic stirrer, holder for pressure transducer, jacketed buffer reservoir and tube set (0.5 L for mouse, 1.0 L for rat/guinea pig

Optional Additions for Accurate Control of Perfusion Pressure or Flow

Addition for perfusion pressure measurement to PSCI: Add 73-1523 PLUGSYS Minicase, Type 609; 73-1793 TAM-D; 73-3862 APT300

Addition for constant pressure perfusion to PSCI: In addition to above, add 73-2806, Servo Controller for Perfusion (SCP)

Setup & Operation

The PSCI works such that individual cells are released from the cell structure of the tissue through perfusion with enzyme solution and are then flushed out. Those cardiomyocytes are collected in the collagenase reservoir to which the dissected left ventricle is added for further dissociation, allowing an increased total cell yield.

The apparatus is so designed that the individual steps required for preparing the cells can proceed as simply and clearly as possible. With two separate perfusion circuits, the organ can be switched between clearing buffer for removal of  blood cells and collagenase buffer for organ digestion by means of a custom, precision-milled changeover stopcock.

Using the standard configuration, perfusion takes place under constant-flow conditions which are set by the researcher on the peristaltic pump. The limits of the apparatus are a flow rate of about 50 or 100 ml/min depending on the configuration (mouse vs. rat/guinea pig).

Common additions to the PSCI include perfusion pressure measurement and constant pressure perfusion. These options allow for optimized perfusion of the heart, further maximizing cell yield and viability by ensuring non-damaging physiological perfusion conditions. The components wetted by the perfusion solutions are made from alcohol-resistant materials so that the apparatus can be filled with ethanol for sterilization/disinfection should your experiment require prolonged incubation time where bacterial contamination would be problematic.

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