Human Placenta Laminin-111 as a Multifunctional Protein for Tissue Engineering and Regenerative Medicine

Adv Exp Med Biol. 2018:1077:3-17. doi: 10.1007/978-981-13-0947-2_1.

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.

Keywords: Laminin-111; NG 108-15; Placenta; Schwann cells; Vasculogenesis.

MeSH terms

  • Cell Line
  • Female
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Laminin / isolation & purification
  • Laminin / metabolism*
  • Placenta / metabolism*
  • Pregnancy
  • Regenerative Medicine*
  • Schwann Cells
  • Tissue Engineering*

Substances

  • Laminin