Directions for Use - Advanced BioMatrix

AlignCol Matrix
Aligned-Braided Collagen Fibrils
on Glass Slides, Sterile
Catalog Number 5054
Product Description
Collagen is the primary structural element in extracellular
matrices (ECM). Type I collagen is the major structural
component of skin, bone, tendon, and other fibrous
connective tissues. Most in vivo tissues demonstrate
structural alignment of collagen fibrils. These aligned
collagen fibrils can provide a mechanically suitable and
information-rich scaffold for cell-ECM interactions.
The application of aligned fibrils of Type I collagen as a
scaffold matrix is a highly attractive prospect. It has the
advantage that cells can be directly influenced by the aligned
collagen fibrils as they grow into an oriented, in vivo-like
network.
Through a process termed “contact guidance,” the cells align
with the fibrils as the fibrils direct their migration. Such
alignment is characteristic of many tissues, and may provide
microstructural and mechanical cues for regulation of cell
phenotype and function, as well as influence the gross
mechanical properties of the tissues.
AlignCol is an engineered scaffold consisting of aligned
Type I collagen fibrils. Using Fibralign Nanoweave
collagen-based materials, the purified collagen is printed
onto borosilicate glass chips at a thickness of ~ 1 micrometer
while aligning the collagen fibrils in an exact orientation. As
the aligned fibrils are applied to a glass slide, the process is
controlled for parameters such as fibril positioning,
mechanical strength, structural uniformity, average fibril sizes
and optical characteristics.
AlignCol Matrix, Aligned-Braided product is provided on glass
slides that measure 8 X 15 mm. An indicator is placed on the glass
slide indicating the side on which the aligned collagen matrix is
provided. Direction for Use provides instruction on the handling of
the product.
Characterization of Type I Collagen Used
Purity: PureCol ® is ultrapure collagen (~99.9% SDS-PAGE, ~97%
Type I with remainder Type III collagen). SDS-PAGE shows the
typical , and banding pattern.
Fibrillogenesis: The formation and development of fibrils present
in collagen fibers are tested and results are characteristic to
standards.
pH: pH ~2.0
Endotoxin: < 0.5 EU/ml (LAL assay)
Product Characterization
Atomic Force Microscopy (AFM): Topology and alignment of fibrils
is confirmed and characteristic as at the picture below.
The Type I collagen used as the starting material for these
aligned collagen fibril matrices is PureCol®, a highly purified
and characterized collagen material.
AlignCol Matrix, Aligned-Braided Collagen product provides
collagen fibril orientation where the fibrils are aligned in the same
direction with a lateral orientation. This configuration replicates
the organization of collagen fibrils in corneal stroma and some
other connective tissues.
Copyright, 2010 Advanced BioMatrix, Inc. Revision 3

Storage: This product is stored and shipped at ambient or room
temperature.
Stability: The product shelf life is 24 months.
Cell Attachment and Growth Assay: To demonstrate cell
attachment and alignment, cells are seeded on the aligned matrix
and meet criteria.
Precautions and Disclaimer
This product is for R&D use only and is not intended for human or
other uses. Please consult the Material Safety Data Sheet for
information regarding hazards and safe handling practices.
Procedure
1. AlignCol Matrix is supplied sterile in a container. Using sterile
forceps, remove the AlignCol Matrix slide from the container by
gripping the end of the slide ensuring that the forceps do not
disturb the matrix portion of the slide.
2. Upon removal of the slide, check for the right-side up
orientation of the matrix. An indicator is placed on the glass slide
indicating the side on which the aligned collagen matrix is
provided.
3. Dip AlignCol Matrix slide into DPBS without Ca and Mg for
5 minutes and remove.
4. Immediately dip and rinse AlignCol Matrix slide into de-ionized
water using a gentle motion for 5-10 seconds and remove.
5. Immediately immerse AlignCol Matrix slide into sterile 70%
ethanol for minimum of 1 hour to sterilize and remove.
6. Place the AlignCol Matrix slide in a new culture dish and let it
dry. Positioning the slide at an angle against the wall (not flat on
the bottom) allows for the ethanol to evaporate faster. Once
dried, place the slide flat on the bottom of the culture dish with
the orientation of the matrix in the up position. Alternatively,
rinse the e slide 3 times with PBS or DMEM (e.g., 3 ml for one well
of 6-well plate) ensuring that the orientation of the matrix is in the up
position.
8. Remove DMEM and add cell suspension in DMEM without
serum into the culture system.
9. After cells attach, remove the DMEM and replace with complete
cell culture medium (e.g. DMEM with serum).
List of References
1. E. Vrana, N. Builles, M. Hindie, O. Damour, A. Aydinli, V. Hasirci, "Contact
guidance enhances the quality of a tissue engineered corneal stroma",
Journal of Biomedical Materials Research Part A, 84, Issue 2 , 2008, 454 –
463.
2. L. Besseaua, B. Coulombb, C. Lebreton-Decosterb, M.-M. Giraud-Guillec,
"Production of ordered collagen matrices for three-dimensional cell
culture", Biomaterials, 23, 2002, 27–36.
3. Cheng Guo, Laura J. Kaufman, "Flow and magnetic field induced collagen
alignment", Biomaterials, 28, 2007, 1105–1114.
4. C. Helarya, L. Ovtrachta, B. Coulombb, G. Godeaub, M. M. Giraud-Guille,
"Dense fibrillar collagen matrices: A model to study myofibroblast
behaviour during wound healing", Biomaterials, 27, 2006, 4443–4452.
5. P. P. Provenzano, K. W. Eliceiri, J.M. Campbell, D. R. Inman, J. G. White,
and P. J. Keely, "Collagen reorganization at the tumor-stromal interface
facilitates local invasion", BMC Medicine, 4:38, 2006.
6. K. Yoshizato, T. Obinata, H.-Yh Huang, R. Matsuda, N. Shioya, and T.
Miyata, "In Vitvo Orientation of Fibroblasts and Myoblasts on Aligned
Collagen Film", Develop., Growth and Differ., 23 (2), 1981, 175-184.
7. H. J. Evans, J. K. Sweet, R. L. Price, M. Yost, and R. L. Goodwin, "Novel 3D
culture system for study of cardiac myocyte development", Am J Physiol
Heart Circ Physiol 285, 2003, H570–H578.
8. N. Dubey, P. C. Letourneaub, R. T. Tranquillo, "Neuronal contact guidance
in magnetically aligned fibrin gels: effect of variation in gel mechanostructural
properties", Biomaterials, 22, 2001, 1065-1075.
9. B. I. Rosner, T. Hang, R.T. Tranquillo, "Schwann cell
behavior in three-dimensional collagen gels: evidence for differential
mechano-transduction and the influence of TGF-beta 1 in morphological
polarization and differentiation", Exp Neurol, 195, 2005, 81-91.
10. H. Haga, C. Irahara, R. Kobayashi, T. Nakagaki, and K. Kawabata,
"Collective Movement of Epithelial Cells on a Collagen Gel Substrate",
Biophysical Journal, 88, 2005, 2250–2256.
11. Lee P, Lin R, Moon J, Lee L P., "Microfluidic alignment of collagen fibers
for in vitro cell culture", Biomed Microdevices, 8, 2006, 35–41.
12. Y.-R. V. Shih, C.-N. Chen, S.-W. Tsai, Y. J. Wang, O. K. Lee, "Growth of
Mesenchymal Stem Cells on Electrospun Type I Collagen Nanofibers",
Stem Cells, 24, 2006, 2391-2397.
13. G. G. Fuller, Langmuir-Blodgett films of oriented collagen films as cell
culture substrates, 81st ACS Colloid & Surface Science Symposium, June
2007.
7. Pre-incubate the slide in DMEM for 45 minutes at 37°C in a CO 2
incubator.
Copyright, 2010 Advanced BioMatrix, Inc. Revision 3