Magnetic separation of human podocalyxin-like protein 1 (hPCLP1)-positive cells from peripheral blood and umbilical cord blood using anti-hPCLP1 monoclonal antibody and protein A expressed on bacterial magnetic particles.
Hemangioblasts are common progenitors of hematopoietic and angiogenic cells, which have been demonstrated in the mouse to possess a unique cell surface marker, podocalyxin-like protein 1 (PCLP1) (Hara, T. et al., Immunity, 11: 567-578. 1999). In this study, we prepared a novel monoclonal antibody against human PCLP1 (hPCLP1) and attempted to isolate human hematopoietic progenitor cells from umbilical cord blood and peripheral blood using nano-sized bacterial magnetic particles (BacMPs) coupled with the anti-hPCLP1 antibody. Flow cytometric analysis demonstrated that the purity of separated hPCLP1-positive cells from peripheral blood was approximately 95% whereas peripheral blood mononuclear cells contained only 0.1% PCLP1+ cells. Umbilical cord blood was demonstrated to be a better source for PCLP1+ cells than peripheral blood. These results suggest that the separation of human PCLP1+ cells using BacMPs with anti-hPCLP1 were extremely effective and may be useful as a means to prepare human hematopoietic progenitor cells.
Kuhara
Motoki
M
Technology and Development Division, Medical & Biological Laboratories Co. Ltd., 1063-103, Ohara, Terasawaoka, Ina, Nagano, 396-0002, Japan. kuhara@mbl.co.jp
Yoshino
Tomoko
T
Shiokawa
Miho
M
Okabe
Tomoya
T
Mizoguchi
Shinji
S
Yabuhara
Akihiko
A
Takeyama
Haruko
H
Matsunaga
Tadashi
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002460:Cell Line; D005312:Fetal Blood / Q000737:chemistry; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology / Q000276:immunology*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D042201:Magnetospirillum / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry; D012795:Sialoglycoproteins / Q000097:blood* / Q000276:immunology* / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000276:immunology
D000911:Antibodies, Monoclonal; D012795:Sialoglycoproteins; D013205:Staphylococcal Protein A; C001643:podocalyxin