1. Human Placenta Laminin-111 as a Multifunctional Protein for Tissue Engineering and Regenerative Medicine
- Author
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Johannes, Hackethal, Christina M A P, Schuh, Alexandra, Hofer, Barbara, Meixner, Simone, Hennerbichler, Heinz, Redl, and Andreas H, Teuschl
- Subjects
Tissue Engineering ,Pregnancy ,Placenta ,Human Umbilical Vein Endothelial Cells ,Humans ,Female ,Laminin ,Schwann Cells ,Regenerative Medicine ,Cell Line - Abstract
Laminins are major components of all basement membranes surrounding nerve or vascular tissues. In particular laminin-111, the prototype of the family, facilitates a large spectrum of fundamental cellular responses in all eukaryotic cells. Laminin-111 is a biomaterial frequently used in research, however it is primarily isolated from non-human origin or produced with time-intensive recombinant techniques at low yield.Here, we describe an effective method for isolating laminin-111 from human placenta, a clinical waste material, for various tissue engineering applications. By extraction with Tris-NaCl buffer combined with non-protein-denaturation ammonium sulfate precipitation and rapid tangential flow filtration steps, we could effectively isolate native laminin-111 within only 4 days. The resulting material was biochemically characterized using a combination of dot blot, SDS-PAGE, Western blot and HPLC-based amino acid analysis. Cytocompatibility studies demonstrated that the isolated laminin-111 promotes rapid and efficient adhesion of primary Schwann cells. In addition, the bioactivity of the isolated laminin-111 was demonstrated by (a) using the material as a substrate for outgrowth of NG 108-15 neuronal cell lines and (b) promoting the formation of interconnected vascular networks by GFP-expressing human umbilical vein endothelial cells.In summary, the isolation procedure of laminin-111 as described here from human placenta tissue, fulfills many demands for various tissue engineering and regenerative medicine approaches and therefore may represent a human alternative to various classically used xenogenic standard materials.
- Published
- 2018