3D Collagen Culture Kit

3D Collagen Culture Kit

Chemicon (Millipore)

Cell migration is a fundamental function of normal cellular processes, including cell proliferation, and migration. Cell behavior in vitro is typically examined in a 2D environment; however, in the body, cells exist in a three-dimensional extracellular matrix environment rich in type I collagen. In fact, cells cultured in 3D matrices better reflect in vivo cell physiology when compared to traditional 2D systems. CHEMICON has developed a three-dimensional cell culture system that closely simulates the collagen-rich extracellular matrix of normal tissue. This 3D system provides a simple and rapid method to analyze cell angiogenesis, migration, apoptosis, proliferation and tissue formation in a 3D-collagen matrix. Cells suspended in this 3D system are easily visualized by phase contrast or fluorescence light microscopy. Cells can be directly fixed and stained within the matrix and treated with antibodies for visualization of specific intra- and extracellular proteins (Please see Appendix B). Importantly, cells suspended in our 3D system can also be treated with various reagents allowing for extensive screening of biological responses to growth factors and chemical agents. As an additional benefit to investigators, sterile and viable cells may be removed from the 3D Cell Culture System for further experimentation, including FACS or biochemical analysis.



ECM675 uses purified chicken type I collagen in 6mM acetate buffer pH 7.0. The product is 99.9% pure native atelomeric avian collagen. Approximately 85% type I and 15% type III collagen protein. The material has been specially prepared to maintain its native state as much as possible. The telomeric ends of the collagen I have been enzymatically removed. Since most of the species identity is found within the telomeric ends, our atelomeric chicken collagen should provide a suitable substrate for most any animal species. The working concentration and precise purification protocol are proprietary and will not be disclosed, however it at least a 0.3% solution of treated avian collagen.

For Research Use Only; Not for use in diagnostic procedures

Activity Assay

The collagen in ECM675 is composed of ~90% chicken collagen type 1 and 10% type III. It is prepared by a novel method that cannot be disclosed completely. There is no enzyme used to strip the product from the tissue. The whole process from start to finish is done below 25C and mostly below 10C. Chicken feet (paw ends only) are washed down to pyrogen free status then minced to 4mm pieces, then soaked in 1M acetic acid. It is this which releases the collagen since the paws are around 75% collagen. The only denaturation done is that an enzyme is used to remove the telomeric ends of the molecules thus making them atelo--the product is therefore best described as atelo collagen. It has no residual chicken identity since much of the species identity is contained in those telomeric ends.

Human

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Collagens are proteins in the Extracellular Matrix (ECM), critical in cell adhesion. At least 17 types of Collagen have been identified, with many genes encoding distinct subunits. Collagen type I is found in several tissues including skin, connective tissue, cartilage and bone. Collagen type II is also found in cartilage, as well as in the vitreous humor of the eye. Collagen type III is found in connective tissue, often in association with type I Collagen. These three Collagens, known as fibrillar or interstitial Collagens, are trimers with a triple helical structure; helices assemble into rods and fibrils held together by inter-trimer crosslinks. Disruptions in expression of these Collagens result in several diseases, such as Ehlers-Danlos syndrome (type I and III), osteoporosis (type I), achondrogenesis or chondrodysplasia (type II), osteoarthritis (type II), or aneurysms (type III). Introduction of antibodies against Collagen type II into mice provide a model for rheumatoid arthritis.

· Tissue Culture Dishes

· Pipettes and Sterile Tips

· Microcentrifuge Tubes, Sterile

· 37oC Incubator

· Inverted Light Microscope

· Bacterial Collagenase

• Collagen Solution - (Part No. 90135) One bottle - 10 mL
• 5X RPMI Medium- (Part No. 90134) One bottle - 2.5 mL
• 5X M199 Medium- (Part No. 90133) One bottle - 2.5 mL
• 5X DMEM Medium- (Part No. 90137) One bottle - 2.5 mL
• 5X PBS with Phenol Red- (Part No. 90136) One bottle - 2.5 mL
• Neutralization Solution- (Part No. 90138) One vial - 0.5 mL

Collagen solution is supplied in 6mM acetate buffer at ~ pH7.0.

Cell Based Assays

Cell Based Assays

Store kit materials at 4°C up to their expiration date.