1 The potential of human hepatic stem cells (hHpSCs) and other stem/progenitors for pharmaceutical research, cell-based therapies, and tissue engineering relies on being able to isolate them, propagate them in culture and differentiate them to a functional mature cell fate(s).2 Current methods for differentiation of stem cells

involve subjecting cells to a mix of soluble signals and/or extracellular matrix components, and the stem cells must be treated with multiple sets of such signals over weeks of time. The Everolimus adult fate achieved is typical of only partially differentiated cells with over- or underexpression of specific adult genes.3 Here we demonstrate strategies for rapidly differentiating stem cells using matrix scaffolds that elicit more efficient and reproducible responses. Extracellular matrix is an extraordinarily complex mixture of molecules that are highly regulated, secreted by, and adjacent to cells on one or more of their surfaces, and long understood to be critical for determining GSK458 the morphology,

growth, and differentiation of attached cells.4, 5 Tissue-specific gene expression in cultured cells is improved by culturing the cells on or embedded in matrix extracts or purified matrix components.6, 7 However, individual matrix components, alone or in combination, are unable to recapitulate a tissue’s complex matrix chemistry and architecture. This is related to the fact that the matrix components are in patterns associated with natural tissue zones and with histological structures such as blood vessels. This complexity of the tissue matrix is more readily achieved by matrix extracts of decellularized

tissue.8-10 Matrix extracts selleck screening library found useful for ex vivo maintenance of cells include amniotic membrane extracts11; Matrigel, an urea extract of a murine embryonal carcinoma12; extracellular matrix (ECM), a detergent- or NaOH-extract of monolayer cell cultures13,14; and biomatrices, an extract of homogenized tissues.10, 15 More recently, decellularized tissues, prepared by collagenase digestion of a tissue16 or by delipidation followed by distilled water washes,8 have been used to mimic the matrix environment in vivo.17 Even though these protocols result in major losses of some matrix components, the decellularized scaffolds from different tissues or organs, such as small intestinal submucosa (SIS), bladder submucosa matrix (BSM),17, 18 vascular tissue,19 heart,20 airway,21 and liver22 have been used successfully in both preclinical and clinical applications.23 Here we describe a strategy, focused on collagen chemistry, that is ideal for preparing substrata of tissue extracts comprised of tissue-specific matrix components and factors bound to the matrix.