title: Phospholipid polymer-based antibody immobilization for cell rolling surfaces in stem cell purification system.
abstract: We previously developed an antibody-conjugated cell rolling column that successfully separates stem cell subpopulations depending on the cell surface marker density, but a large amount of the injected cells were retained in the column because of non-specific interactions. In this study, an amphiphilic copolymer, poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (nBMA)-co-N-vinyl formamide (NVf)], with phospholipid polar side groups was designed as a novel antibody-immobilizing modifier. The formamide groups in NVf units were converted to active maleimide groups. A plastic flow microfluidic chamber was coated with the copolymers, and a reduced anti-CD90 antibody was immobilized. The adipose tissue-derived stem cells isolated from the rat were injected into the flow chamber, and their rolling behavior was observed under a microscope with a high-speed camera. Non-specific cell adhesion was reduced strongly by means of this immobilization method because of the MPC unit, resulting in a high percentage of rolling cells. These results demonstrate that a surface coated with phospholipid polar groups can be used in an effective stem cell separation system based on the cell rolling process.
authors.affiliation: a Department of Biomedical Engineering , National Cerebral and Cardiovascular Center Research Institute , Suita , Osaka 565-8565 , Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Mahara
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Ishihara
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000327:Adsorption; D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry*; D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D002469:Cell Separation; D057927:Hydrophobic and Hydrophilic Interactions; D008297:Male; D008689:Methacrylates / Q000737:chemistry; D046208:Microchip Analytical Procedures; D010743:Phospholipids / Q000737:chemistry* / Q000494:pharmacology*; D051381:Rats; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects*; D013499:Surface Properties
chemical_list: D055910:Antibodies, Immobilized; D001672:Biocompatible Materials; D008689:Methacrylates; D010743:Phospholipids; C016284:butyl methacrylate
keywords: 2-methacryloyloxyethyl phosphorylcholine; cell rolling; microfluidic camber; stem cell