Fig. 3. Growth and differentiation of sorted ALB^– and ALB⁺ cells cultured with GFs and ECMs. FACS-sorted ALB^– and ALB⁺ cells were cultured separately. For the GF test, cells were cultured with or without HGF, OSM, aFGF, bFGF or a mixture of all GFs on type IV collagen-coated dishes. For the ECM test, cells were cultured on either non-coated, or laminin-, type IV collagen-, type I collagenor fibronectin-coated dishes. After 10 days in culture, proliferation (A) and differentiation (B) of sorted cells were examined by cell counting (A) or FACS (B). Representative data from a transfected stem cell clone are shown; three samples were examined for each GF or ECM. Data are mean±s.d. (A) HGF strongly promoted the proliferation of both ALB^– and ALB⁺ sorted cells, whereas lesser effects were noted with OSM, aFGF and bFGF. OSM specifically suppressed the proliferation of ALB^– cells. For the ECMs, laminin, type IV collagen and type I collagen induced the proliferation of ALB⁺ cells, but to a lesser degree than HGF. All data were normalized to the value of no-GF (GF test) or a non-coated dish (ECM test) and fold-differences are shown. (B) FACS-analysis revealed that HGF and OSM have the potential to induce ALB⁺ cells from ALB^– sorted cells, and inhibit generation of ALB^– cells from ALB⁺ sorted cells. Laminin, type IV collagen and type I collagen possessed similar, but lesser, effects than did HGF and OSM. All data are normalized to the value at the day of sorting (day 0) (GF test) or to the value of a non-coated dish (ECM test) and fold differences are shown.

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