A fusion protein N-cadherin-Fc as an artificial extracellular matrix surface for maintenance of stem cell features.
N-cadherin is a cell-cell adhesion molecule and plays important roles in neural development. With conventionally used extracellular matrices (ECMs), maintenance of undifferentiated state of stem cells and regulation of their neural differentiation process is very difficult due to the colony formation through intercellular interactions. To overcome the above-mentioned problems, we developed a new artificial ECM to mimic N-cadherin-mediated cell adhesion. In this study, we constructed a chimeric protein (N-cadherin fused to IgG-Fc, abbreviated as N-cad-Fc), which contains extracellular domain of N-cadherin and Fc domain of IgG. We confirmed that N-cad-Fc can stably adsorb to hydrophobic surface. We checked maintenance of undifferentiated state and neural differentiation ability of stem cells cultured on N-cad-Fc-coated surface. Both P19 and MEB5 cells cultured on N-cad-Fc-coated surface showed scattering morphologies without colony formation and higher proliferating potency than conventional culture systems with maintenance of undifferentiated state. Both of two cell lines cultured on N-cad-Fc-coated surface differentiated into neural cells at a single cell level when induced with proper conditions. Furthermore, the expression of neuron-related gene Neurog1 in two cell lines cultured on N-cad-Fc-coated surface was promoted. Therefore, it will be expected that the constructed N-cad-Fc can be used as an artificial ECM for stem cells.
Yue
Xiao-Shan
XS
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-57, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Murakami
Yuta
Y
Tamai
Toshiyuki
T
Nagaoka
Masato
M
Cho
Chong-Su
CS
Ito
Yoshihiro
Y
Akaike
Toshihiro
T
D000818:Animals; D015820:Cadherins / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011961:Receptors, Fc / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects*; D013499:Surface Properties / Q000187:drug effects
D015820:Cadherins; D011961:Receptors, Fc; D011993:Recombinant Fusion Proteins