Biofunctional Thermo-Responsive Polymeric Surface with Micropatterns for Label Free Cell Separation
- PDF / 1,139,291 Bytes
- 6 Pages / 432 x 648 pts Page_size
- 50 Downloads / 139 Views
Biofunctional Thermo-Responsive Polymeric Surface with Micropatterns for Label Free Cell Separation Yoshikazu Kumashiro1, Jun Ishihara1,2, Terumasa Umemoto1, Kazuyoshi Itoga1, Jun Kobayashi1, Masayuki Yamato1 and Teruo Okano1 1 Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku,Tokyo 162-8666, Japan 2 Division of Cellular Therapy, The Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8039, Japan. ABSTRACT Thready stripe-patterned thermo-responsive surfaces were prepared and their surface properties were characterized. Prepared 3 μm wide stripe-patterned surfaces were evaluated by observing the adhesions and detachments of three types of cells: HeLa cells (HeLas), human umbilical vein endothelial cells (HUVECs), and NIH-3T3 cells (3T3s). Although cell adhesion and detachment in response to temperature were observed on all cells on a conventional thermoresponsive surface without patterns, the thermo-responsive surface with a 3 μm striped-pattern exhibited various cell adhesion properties. HeLas hardly adhered to the patterned surface even at 37 ºC. On the other hand, although HUVECs adhered on the patterned surface at 12 h after incubation at 37 ºC, the adhered HUVECs detached themselves after another 12 h incubation at 37 ºC. 3T3s adhered to the patterned surface at 37 ºC and detached themselves after reducing temperature to 20 ºC. A mixture of HeLa, HUVEC and 3T3 was separated using their different specific cell-adhesion properties, and the composition of cells was analyzed by a flow-cytometry. As a result, the conventional thermo-responsive surface with a stripe-pattern was found to function as a cell-separating interface by using specific cell adhesion properties. INTRODUCTION Functional surfaces have attracted much attention through their induction of various potential applications, such as superhydrophobic and superhydrophilic surfaces inspiring the nanostructure of lotus leaf [1,2], stimuli-responsive surfaces in response to external stimuli (pH [3,4], temperature [5], ultra-violet light [6,7], and so on), and self-governing surfaces [8,9]. In our laboratory, thermo-responsive cell culture dishes having poly(N-isopropylacrylamide) (PIPAAm) covalently immobilized on tissue culture polystyrene, are used for preparing an artificial tissue consisting of two dimensional monolayer cells [10-13]. Cells adhere and proliferate at 37 ºC on PIPAAm modified cell culture surface, because PIPAAms are hydrophobic owing to their dehydration. After reducing temperature to 20 ºC, the adhered cells detach themselves as single cells or a contiguous cell sheet from the surfaces, due to the hydration and swelling of grafted PIPAAms on the culture surface. Cell therapies including tissue engineering and cell injection have focused in the field of regenerative medicine that reproduces the lost functions of the tissue and organs, and some clinical trials by transplantation of cells have been already performed for recovering th
Data Loading...
