Effects of Extracellular Matrices on F9 Teratocarcinoma Stem Cells: A Crucial Role of Type IV Collagen in the Early Stage of Differentiation of F9 Stem Cells

Abstract

F9 teratocarcinoma stem cells treated with retinoic acid (RA) differentiate into visceral endoderm, and this process affects the expression of some extracellular matrix (ECM) proteins. As the direct influence of ECM molecules on the differentiation of F9 stem cells has not been studied, we investigated the effects of various ECM proteins on the early differentiation of these cells. Monolayers of F9 stem cells were cultured on dishes coated with various ECM, such as type I or IV collagen, fibronectin, or laminin. They aggregated and formed spheroid bodies in the absence of RA only on type IV collagen. The outer layer cells of the spheroid bodies exhibited numerous microvilli, junctional complexes and mature cell organelles. Alpha-fetoprotein was positive in the outer layer cells. A small amount of laminin was detected in the matrix of the spheroid bodies. These data suggest that type IV collagen promoted the early stage of the differentiation of F9 stem cells without RA. The other ECM molecules failed to induce them to form spheroid bodies. The reversibility of the structure of the spheroid bodies to a monolayer was also examined. When the spheroid bodies were reseeded only on fibronectin- or laminin-coated culture dishes, they broke down and the cells spread to the surface in the absence of RA. The differentiation of F9 stem cells induced by type IV collagen seemed to be reversible and to take place at an early stage of their morphogenesis. These findings suggest that type IV collagen plays an important role in early embryogenesis.

Biomimetic Cell Culture Proteins as Extracellular Matrices for Stem Cell Differentiation

“…189 Tables 7 and 8 summarize several types of collagen materials and scaffolds for MSC differentiation that have been reported in the literature. 34,37,40,45,46,53,56,61,63,70,71,79,83,84,91,97,98,101,105,110,141,144,146,148,149,151,154,182−185,189−217 5.2.1. Collagen Type I Scaffolds.…”

“…Typically, 2D cell culture dishes are coated with ECM proteins or ECM-mimicking peptides. Tables and show examples of the ECM proteins and ECM-mimicking peptides used to coat culture dishes and their binding sites on stem cells. ,,,,, ,,− Collagen types I, II, and IV, gelatin, laminin, laminin-1, laminin-5, vitronectin, and fibronectin are typically used as coating materials. ,,,, − ,− ECM-mimicking peptides (e.g., RGD, DGEA, YIGSR, IKVAV, KRSR, P15, and GFOGER) are commonly used as coating or grafting materials. ,,,,− Covalent binding is preferable for long-term effects in culture, but noncovalent coating is the simplest method for the preparation of dishes with immobilized ECM proteins or ECM-mimicking peptides. Figure summarizes typical surface reactions for the covalent immobilization of ECM proteins and peptides on dishes.…”

“…Once the gel has set, extra culture medium is added to the top of the gels and the cultures are returned to the incubator . Tables and summarize several types of collagen materials and scaffolds for MSC differentiation that have been reported in the literature. ,,,,,,,,,,,,,,,,,,,, ,,,,­,,,− ,− …”