Neonatal Cardiomyocyte Isolation System Product Information

The Worthington Neonatal Cardiomyocyte Isolation System has been developed to provide researchers with a reliable, convenient, and consistent neonatal rat cardiomyocyte cell isolation method. By using purified rather than crude enzyme preparations, it has been possible to minimize the lot-to-lot variation. In addition, Worthington use-tests the kits by isolating cardiomyocytes from neonatal rat hearts to assure performance, reliability and consistent yield of viable cells.

The kit has been formulated in collaboration with Dr. Ronal MacGregor. The method is based on that described by Toraason, et al. Toxicol, 56, 107 (1988) in which the minced tissue is incubated overnight with purified trypsin at 2-8°C. As pointed out by Toraason, this step reduces the hands-on time required to harvest cells compared to the time involved in sequential incubations in warm trypsin or collagenase. Purified collagenase rather than crude collagenase is used to maximize yield and viability.

Contents of Kit
The package contains sufficient materials for five separate tissue dissociations, each containing up to twelve hearts. For larger or smaller tissue samples prepare proportionate volumes of reagents at each step and combine them in the same ratio as described in the protocol.

• Vial 1: 1 bottle, 500ml: Sterile calcium- and magnesium-free Hank's Balanced Salt Solution (CMF HBSS), pH 7.4. The solution is used for reconstituting the contents of Vials #2 and #3 in addition to serving as the medium for the dissociation.

• Vial 2: 5 vials, 1000µg each: Worthington Trypsin (Code: TRLS), 3X crystallized, dialyzed against 1mM HCl, filtered through 0.22 micron pore size membrane, and lyophilized. Before use, reconstitute with 2ml CMF HBSS (Vial #1) and swirl gently to dissolve contents. Store at 2-8°C.

• Vial 3: 5 vials, 2000µg each: Worthington Soybean Trypsin Inhibitor (Code: SIC), a 0.22 micron pore size membrane filtered, lyophilized powder. Before use, reconstitute with 1ml CMF HBSS (Vial #1) and swirl gently to dissolve contents. Store at 2-8°C.

• Vial 4: 5 vials, 1500 Units each: Worthington Purified Collagenase (Code: CLSPA), a 0.22 micron pore size membrane filtered, lyophilized powder which has been chromatographically purified. It contains less than 50 caseinase units per milligram and is composed of two separable but very similar collagenases. Before use, reconstitute with 5ml Leibovitz L-15 media (prepared as described below) and swirl gently to dissolve contents. Store at 2-8°C.

• Pouch containing Leibovitz L-15 Media Powder: 1 x 1L, Reconstitute entire contents of pouch by cutting open top of envelope and pouring contents into beaker containing 800ml of cell culture grade water. Rinse pouch 2-3 times with additional 100ml. Bring total volume to 1 liter and filter through a 0.22 micron pore size filter.

The kit also includes 5 Cell Strainers (Falcon), a card correlating phenol red color with pH for checking balanced salt solutions and culture media.

Related Products:

Collagenase (CLS1-4/CLSPA)
Deoxyribonuclease I (DP/D/DCLS/D2/DPFF/DPRF)
Hyaluronidase (HSE/HSEP)
Neutral Protease (Dispase, NPRO)
Papain (PAP/PAPL/PAP2)
Proteinase K (PROK)
Trypsin (TL/TRL/TRL3/TRLS/TRTPCK)Trypsin Inhibitors (LBI/OI/SI/SIC)

Cell Isolation Optimizimg System (CIT)
Hepatocyte Isolation System (HIS)
Papain Dissociiation System (PDS/PDS2)

DescriptionActivityCodeSizeCat. No.Price
Neonatal Cardiomyocyte Isolation System
Kit for performing five separate tissue dissociations each containing up to twelve hearts. Contains single use vials of purified collagenase and trypsin, CMF-HBSS, Leibovitz L-15 media and Falcon cell strainers along with a detailed protocol. The kit is use-tested by Worthington to assure performance.
Store at 2-8C.
NCIS 1 ki LK003300 $228.00 Buy Now
3 ki LK003303 $595.00 Buy Now
Collagenase Vial, NCIS
Source: Clostridium histolyticum
A component of the NCIS kit. This material is 0.22 micron membrane filtered and lyophilized in autoclaved vials. A vial reconstituted with 5 ml of HBSS or equivalent yields a solution of 300 units/ml of collagenase, Code: CLSPA. Suitable for cell isolation and culture applications.
Store at 2-8C.
≥500 units per mg dry weight CLSPANK 1 vi LK003240 $28.00 Buy Now
5 vi LK003245 $110.00 Buy Now
Trypsin Vial, NCIS
Source: Bovine Pancreas
A component of the NCIS kit. This material is 0.22 micron membrane filtered and lyophilized in autoclaved vials. A vial reconstituted with 2 ml of HBSS yields a solution of 500µg/ml of trypsin, Code: TRLS. Suitable for cell isolation and culture applications.
Store at 2-8C.
≥180 units per vial TRLSNK 1 vi LK003220 $11.00 Buy Now
5 vi LK003225 $40.00 Buy Now
Inhibitor Vial, NCIS
Source: Soybean
A component of the NCIS kit. This material is 0.22 micron membrane filtered and lyophilized in autoclaved vials. A vial reconstituted with 1 ml of HBSS or equivalent yields a solution of 2 mg/ml of trypsin inhibitor, Code: SIC. Suitable for cell isolation and culture applications.
Store at 2-8C.
N/A SICNK 1 vi LK003230 $12.00 Buy Now
5 vi LK003235 $38.00 Buy Now
HBSS Solution
Sterile calcium and magnesium free Hank's balanced salt solution (CMFHBSS), pH 7.4, as supplied in the NCIS kit; 1 x 500 ml.
Store at 2-8C.
HBSS 1 ea LK003210 $52.00 Buy Now
L-15 Media Powder (L15NK)
Leibovitz L-15 media powder, a component of the NCIS kit. Reconstitute entire contents of pouch, QS to 1 liter with cell culture grade water, and 0.22 micron filter. Suitable for cell isolation and culture applications.
Store at 2-8C.
L15NK 1 ea LK003250 $24.00 Buy Now
Cell Strainers (Falcon)
Cell strainers (Falcon), components of the NCIS kit. Suitable for removal of tissue debris in cell isolation applications.
Store at room temperature.
CELSTRNK 5 ea LK003265 $24.00 Buy Now

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