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
发布: 2012年11月05日第2卷第21期 DOI: 10.21769/BioProtoc.281 浏览次数: 16757
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参见作者原研究论文
The authors used this protocol in:
Mar 2012
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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
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
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.
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分类
干细胞 > 成体干细胞 > 造血干细胞
细胞生物学 > 细胞分离和培养 > 细胞分离
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