Enhanced reseeding of decellularized rodent lungs with mouse embryonic stem cells

Lecht S., Stabler C.T., Rylander A.L., Chiaverelli R., Schulman E.S., Marcinkiewicz C., Lelkes P.I.Biomaterials, 2014

Pre-treatment of the decellularized lung with defined ECM proteins, to evaluate the efficacy of reseeding of mESC. All current decellularization methodologies result in significant alterations of the contents, and ratios of ECM proteins, though the exact degree of the loss of ECM glycoproteins, such as laminins. Following ECMs were tested: bovine collagen type IV; human collagen type IV; human pro-collagen type I; human collagen type I; rat collagen type I; human plasma-purified fibronectin; human thrombospondin-1; VCAM; vitronectin; bovine elastin; Matrigel-purified laminin 111; and human recombinant laminins 111, 211, 332, 411, 421, 511, 521. mESCs lack major integrin receptors for collagens but express high levels of functional receptors for LM and FN. Laminin next to FN appears to be the major pro-adhesive ECM protein for mESCs. The mESCs were found to adhere differentially in an isoform-dependent manner with the following order of potencies 511 = 521 >332 >421 >211 >111 >411. These findings further support the notion that a3b1 and a6b1 integrin receptors expressed on the mES cell surface are involved in the specific binding to LM in general and to LM 511 and 521. LM and FN interact more efficiently with collagen type I than with collagen IV or elastin 90% of the available Coll I in the decellularized lung matrix forms a complex with LM. in vitro that cell-derived FN and LM interact with Coll I with much higher efficiencies than commercial FN or LM This finding may be explained by a possible partial loss of activity as a result of the purification procedures in contrast to the unprocessed CM.