title: Orientation-regulated immobilization of Jagged1 on glass substrates for ex vivo proliferation of a bone marrow cell population containing hematopoietic stem cells.
abstract: Notch signaling has been recognized as a key pathway to regulate the proliferation and differentiation of hematopoietic stem cells (HSC). In this study, the orientation-regulated immobilization of a Notch ligand was designed to achieve the efficient Notch ligand-receptor recognition for the ex vivo proliferation of a bone marrow cell population containing HSC. Protein A was chemically conjugated onto aminated glass substrates, followed by immobilizing a recombinant chimeric protein of Jagged1 and Fc domain (Jagged1-Fc) through the biospecific binding between protein A and Fc domain. Protein A adsorption was suppressed for the Jagged1-Fc-immobilized substrates, in contrast to the Jagged1-Fc-coated ones, indicating the orientation-regulated immobilization of Jagged1-Fc for the substrates. Mouse lineage negative cells (Lin(-)) were cultured on the Jagged1-Fc-immobilized substrates. Flow cytometric analyses demonstrated that c-Kit(+), Sca-1(+), Lin(-), and CD34(-) cells of an HSC population was significantly proliferated on the Jagged1-Fc-immobilized substrates 6 days after culture, whereas no proliferation was observed for the Jagged1-Fc-coated substrates in a random manner or Jagged1-Fc-immobilized ones with a Notch signaling inhibitor. It is concluded that the orientation-regulated immobilization of Jagged1-Fc increased the efficiency of Jagged1 to recognize the Notch receptors, resulting in the promoted ex vivo proliferation of the HSC population.
authors: Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
authors: Hiroyuki
authors: H
authors: Toda
authors: Masaya
authors: M
authors: Yamamoto
authors: Hiroshi
authors: H
authors: Kohara
authors: Yasuhiko
authors: Y
authors: Tabata
mesh_terms: D000327:Adsorption; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D005898:Glass / Q000737:chemistry*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D000072100:Jagged-1 Protein; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D015394:Molecular Structure; D051880:Receptors, Notch / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D000072099:Serrate-Jagged Proteins; D015398:Signal Transduction / Q000502:physiology; D013205:Staphylococcal Protein A / Q000378:metabolism; D013499:Surface Properties
chemical_list: D002135:Calcium-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; C000606274:Jag1 protein, mouse; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins; D000072099:Serrate-Jagged Proteins; D013205:Staphylococcal Protein A