磁珠法从外周血液中分离人血前体细胞和干细胞

Salma Hasan; Isabelle Plo

doi:10.21769/BioProtoc.281

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Peer-reviewed

Isolation of Human Blood Progenitor and Stem Cells from Peripheral Blood by Magnetic Bead

磁珠法从外周血液中分离人血前体细胞和干细胞

Salma Hasan

Isabelle Plo email

发布: 2012年11月05日第2卷第21期 DOI: 10.21769/BioProtoc.281 浏览次数: 16227

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参见作者原研究论文

The authors used this protocol in:

Cover of Oncogene, featuring study using the protocol.

Mar 2012

实验方案合集

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Abstract

The antigen CD34 is a well-known marker present on human progenitor and stem cells. This protocol explains the isolation of CD34⁺ cells from peripheral blood using magnetic bead separation technique. The approximate abundance of CD34⁺ cells in blood is 0.1% of mononuclear cells.

Keywords: Hematopoiesis (造血)

Progenitors (脑细胞前体)

Isolation (隔离)

Materials and Reagents

CD34⁺ cells
Peripheral blood sample (at least 50 ml)
Dextran solution (Sigma-Aldrich, catalog number: D1037-500G )
PBS (Life Technologies, Invitrogen™, catalog number: 14190-094 )
Ficoll human (PAA Laboratories GmbH, catalog number: P04-60500 )
Fetal calf serum (FCS) (Hyclone, catalog number: SV30160.03 )
CD34 MicroBeads and FcR blocking reagent (Miltenyi Biotec, catalog number: 130-046-702 )
APC mouse anti-human CD34 antibody (BD Biosciences, catalog number: 555824 )
EDTA
2% dextran solution
1 L 2% dextran solution (see Recipes)

Equipment

Centrifuges
Auto MACS Pro separator (Miltenyi Biotec)
30 μm nylon mesh (Miltenyi Biotec, catalog number: 130-041-407 )
Tissue culture hood
Flow cytometer
Vacuum filter unit (22 μm, GP Millipore Express PLUS membrane)

Procedure

English

中文翻译

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如何引用

Hasan, S. and Plo-Azevedo, I. (2012). Isolation of Human Blood Progenitor and Stem Cells from Peripheral Blood by Magnetic Bead . Bio-protocol 2(21): e281. DOI: 10.21769/BioProtoc.281.