Capture and release of cells using a temperature-responsive surface that immobilizes an antibody through DNA duplex formation.
We synthesized a temperature-responsive surface that immobilized an antibody via DNA duplex formation for selective capture and release of target cells. Polyethylene films were modified by grafting poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAAm-co-AAc)), which were prepared at various ratios of NIPAAm/AAc. The increased hydrophilicity of P(NIPAAm-co-PAA) film with decreased temperature was confirmed by water contact angle measurement. Single strand DNA (20mer) was chemically immobilized on the surface and then antibody (anti-mouse CD45, mCD45) modified with the complementary single strand DNA was immobilized on the surface through DNA duplex formation. The mCD45 antibody immobilization was confirmed by immunostaining. HeLa cells (mCD45 negative) and mouse bone marrow (BM) cells (mCD45 positive) were adhered on the surfaces at 37 °C. Although HeLa cells were detached by 4 °C incubation, BM cells were still adhered on the surface and then the adhered cells were released by DNase treatment. From these results, it was suggested that cells could be selectively captured and collected by using a film having surface that immobilizes an antibody via DNA duplex formation.
a Institute of Biomaterials and Bioengineering , Tokyo Medical and Dental University , Tokyo , Japan.
Tsuyoshi
T
Kimura
a Institute of Biomaterials and Bioengineering , Tokyo Medical and Dental University , Tokyo , Japan.
Naoko
N
Nakamura
a Institute of Biomaterials and Bioengineering , Tokyo Medical and Dental University , Tokyo , Japan.
Kanji
K
Umeda
a Institute of Biomaterials and Bioengineering , Tokyo Medical and Dental University , Tokyo , Japan.
Yoshihide
Y
Hashimoto
a Institute of Biomaterials and Bioengineering , Tokyo Medical and Dental University , Tokyo , Japan.
Akio
A
Kishida
D000180:Acrylic Resins / Q000737:chemistry; D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry* / Q000276:immunology; D002448:Cell Adhesion; D004247:DNA / Q000737:chemistry*; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D017493:Leukocyte Common Antigens / Q000276:immunology; D008297:Male; D051379:Mice; D009690:Nucleic Acid Conformation; D020959:Polyethylene / Q000737:chemistry; D013499:Surface Properties; D013696:Temperature
D000180:Acrylic Resins; D055910:Antibodies, Immobilized; C052970:poly-N-isopropylacrylamide; C006903:carbopol 940; D020959:Polyethylene; D004247:DNA; D017493:Leukocyte Common Antigens
Selective capture and release of cell; antibody immobilization; polymer graft; surface modification; temperature-response