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

Abstract

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.