Cell Lines

Glycosan offers two options for General Cell Culture of immortalized cell lines:

  • Extracel®: a general starting point for optimization of a cell’s microenvironment. Extracel contains Gelin-S or thiolated gelatin (denatured collagen) which allows attachment of a wide variety of cell types and takes the guesswork out of the appropriate attachment factors to use. Additionally, extracellular matrix proteins can be incorporated non-covalently before gelation or attachment peptides having an N-terminal cysteine can be covalently linked to the matrix8.
  • Extracel-HP®: for applications requiring slow release of growth factors in a cell’s microenvironment. Extracel-HP contains Heprasil®, which contains small amounts of thiolated heparin. The chemically-modified heparin ionically binds a wide variety of growth factors and slowly releases them over time9,10,11.

The Extracel® Hydrogel Kits (Extracel and Extracel®-HP) emulate a simplified version of the extracellular matrix (ECM). Extracel is composed of Glycosil® (thiol-modified hyaluronan), Gelin-S® (thiol-modified denatured collagen), and Extralink® (thiol-reactive crosslinking agent). Thiol-modified heparin is added to Extracel to produce Extracel-HP. Cells can be encapsulated during crosslinking, where they attach and grow within the hydrogel matrix, or they can be plated on top of the hydrogel for pseudo three-dimensional growth. If cells are capable of forming spheroids (e.g. HepG21,2, HepG2 C3A1,2, MDA-MB-2313) or acini (e.g. MDCK2,4), they will do this when encapsulated.

Additional components such as growth factors (GFs) and ECM proteins can be added to mimic a specific ECM environment. The compliance of the final hydrogel can be altered either by varying the concentrations of the hydrogel solutions or by changing the amount of Extralink used to crosslink the hydrogel. All Extracel Hydrogel Kits are tested for bacterial growth and endotoxins.

The following cell lines have been cultured in Extracel-based hydrogels:

  • HepG2 C3A cells1,2
  • HepG2 cells1,2
  • MDCK cells2,4
  • MCF-10A3
  • L929 fibroblasts5
  • NIH 3T3 fibroblasts6
  • HuVEC7

References

  1. G. D. Prestwich, Y. Liu, M. Serban, B. Yu, X. Z. Shu, and A. Scott, “3-D Culture in Synthetic Extracellular Matrices: New Tissue Models for Drug Toxicology and Cancer Drug Discovery,” invited, Adv. Enz. Res., in press (2007).
  2. Unpublished data from G. D. Prestwich, et al, University of Utah.
  3. Y. Liu, X. Z. Shu, and G. D. Prestwich, “Tumor Engineering: Orthotopic Cancer Models in Mice Using Cell-Loaded, Injectable, Crosslinked Hyaluronan-Derived Hydrogels,” Tissue Engineering 13(5), 1091-1101(2007).
  4. Unpublished data from Yongzhi Qiu, Robert McCall, Vladimir Mironov, Xuejun Wen, Clemson University, and Medical University of South Carolina.
  5. Y. Liu, X. Z. Shu, G. D. Prestwich, “Biocompatibility and Stability of Disulfide-Crosslinked Hyaluronan Films,” Biomaterials, 26, 4737-4746 (2005).
  6. X. Z. Shu, Y. Liu, Y. Luo, M. C. Roberts, and G. D. Prestwich, “Disulfide-Crosslinked Hyaluronan Hydrogels,” Biomacromolecules, 3, 1304-1311 (2002).
  7. Unpublished data from G. D. Prestwich, et al, University of Utah.
  8. X. Z. Shu, K. Ghosh, Y. Liu, F. S. Palumbo, Y. Luo, R. A. Clark, and G. D. Prestwich, “Attachment and spreading of fibroblasts on an RGD peptide-modified injectable hyaluronan hydrogel” J. Biomed. Mat. Res. 68A, 365-375 (2004).
  9. S. Cai, Y. Liu, X. Z. Shu, G. D. Prestwich, “Injectable glycosaminoglycan hydrogels for controlled release of human basic fibroblast growth factor”Biomaterials, 26, 6054-6067 (2005).
  10. D. B. Pike, S. Cai, K. R. Pomraning, M. A. Firpo, R. J. Fisher, X. Z. Shu, G. D. Prestwich, R. A. Peattie, “Heparin-regulated release of growth factors in vitro and angiogenic response in vivo to implanted hyaluronan hydrogels containing VEGF and bFGF” Biomaterials, 27, 5242-5251 (2006).
  11. Unpublished data from G. D. Prestwich, et al, University of Utah, and R. Peattie, et al, University of Oregon.