Laminin-111, a 800 kDa protein, plays a fundamental role during embryogenesis in mammals, including cell adhesion, migration, proliferation and differentiation. Hence, laminin-111 is a frequently used biomaterial in research, however isolated in most cases from animal tumors or with cost-intensive recombinant techniques.
Recently, the Ludwig Boltzmann Institute for Experimental and Clinical Traumatology has established an effective method for isolating laminin-111 from human placenta, a medical waste material mainly composed of fetal elements. By using a few smart and non-toxic isolation steps, native laminin-111 (pLm-111) could be isolated within only four processing days.
In vitro, pLm-111 promotes effective adhesion of primary Schwann cells (Figure 1), outgrowth of NG-108-15 neural cell lines (Figure 2), and spontaneous formation of interconnected vascular networks of HUVEC cells (Figure 3).
In summary, the availability of pLm-111 offers new, fully-human research approaches for Tissue Engineering and Regenerative Medicine and may provide a new platform technology for clinical use with an increased overall safety profile for patients.
Figure 1: Schwann cell viability (MTT assay) and proliferation (BrdU assay) 24 hours after seeding on tissue culture plastic (TCP), EHS laminin-111 and pLm-111 coated wells, as well as poly-L-lysin/EHS laminin-111 and poly-L-lysin/pLm-111 (100 µg/mL; mean ± SD; n=18).
Figure 2: Upper panel: fluorescence micrographs of phalloidin (green) and DAPI (blue) stained NG 108-15 cells cultivated on tissue culture plastic (TCP), EHS laminin-111 or pLm-111 at concentrations of 100 µg/mL; Scale bars = 100 µm; Lower panel: analysis of the neurite outgrowth of NG108 cells after 24, 48 and 72 hours of cultivation. (mean ± SD; n=12).
Figure 3: pLm-111 promotes vasculogenesis using gfpHUVEC (a) Fluorescence micrographs of gfpHUVEC 48 hours after seeding on EHS laminin-111, pLm-111, or EHS collagen-4 at concentrations of 1mg/mL. Scale bar = 400 µm. (b) Network characteristics of gfpHUVEC, seeded on EHS laminin-111 or pLm-111 from two independent donors (pLm-111 D1/D2; mean ± SD; n=12).