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

Motoki Kuhara, Tomoko Yoshino, Miho Shiokawa, Tomoya Okabe, Shinji Mizoguchi, Akihiko Yabuhara, Haruko Takeyama, Tadashi Matsunaga

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)23-30
Number of pages8
JournalCell Structure and Function
Volume34
Issue number1
DOIs
Publication statusPublished - 2009
Externally publishedYes

Fingerprint

Staphylococcal Protein A
Fetal Blood
Blood Cells
Monoclonal Antibodies
Hematopoietic Stem Cells
Hemangioblasts
Proteins
podocalyxin
Immunity
Antibodies

Keywords

  • Bacterial magnetic particles (BacMPs)
  • Cell separation
  • Hematopoietic progenitor cells
  • Podocalyxin-like protein 1 (PCLP1)

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Physiology

Cite this

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. / Kuhara, Motoki; Yoshino, Tomoko; Shiokawa, Miho; Okabe, Tomoya; Mizoguchi, Shinji; Yabuhara, Akihiko; Takeyama, Haruko; Matsunaga, Tadashi.

In: Cell Structure and Function, Vol. 34, No. 1, 2009, p. 23-30.

Research output: Contribution to journalArticle

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AU - Okabe, Tomoya

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AU - Takeyama, Haruko

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