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| Neural Crest Stem Cell MicroBeads |
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| Overview |
| The Neural Crest Stem Cell MicroBeads were developed for the positive selection or depletion of neural crest stem cells differentiated from pluripotent stem cells. A method for in vitro–differentiation of neural crest stem cells from human iPS cells has been published and is available in the download section. |
| Details |
Background information
CD271, also known as LNGFR (low-affinity nerve growth factor receptor), NGFR (nerve growth factor receptor), or p75NTR (neurotrophin receptor), belongs to the tumor necrosis factor receptor superfamily. CD271 was initially described to be expressed on cells of the nervous system and was suggested to be involved in the development, survival and differentiation of neural cells.1 CD271 can be found in the central and peripheral nervous system on autonomic and sensory neurons2 as well as on glial cells, including oligodendrocytes3, astrocytes4, Schwann cells5,6, and neural crest stem cells7. Neural crest stem cells can be induced from pluripotent stem cells by synergistic action of two inhibitors of SMAD signaling, Noggin and SB4315428,9 or alternatively, by a single small molecule dorsomorphin blocking signalling of several (TGF-β) superfamily receptors10. CD271+ neural crest stem cells can be selected ten days post-induction. They show expression of typical neural crest markers (HNK1, AP2) and they can be differentiated to homogenous peripheral neurons11.
The CD271 antibody is reported to cross-react with the CD271 antigen in monkey, goat, dog, pig, and sheep12. |
| Columns |
| For positive selection: MS or autoMACS Columns. For depletion: LD or autoMACS Columns. |
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| Figure 1 |
| Neural crest stem cells were in vitro–differentiated from human iPS cells using dorsomorphin for 10 days (10, 11) and subsequently isolated using Neural Crest Stem Cell MicroBeads, two MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD271 (LNGFR)-PE (# 130-091-885) and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence. |
| Before separation |
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| After separation |
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| Details |
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| References |
| 1. Thomson, T. M. et al. (1988) Exp. Cell Res. 174 (2): 533–539. |
| 2. Kashiba, H. et al. (1995) Brain Res. Mol. Brain Res. 30 (1): 158–164. |
| 3. Casaccia-Bonnefil, P. et al. (1996) Nature 383 (6602): 716–719. |
| 4. Rudge, J. S. et al. (1994) Eur J. Neurosci. 6 (5): 693–705. |
| 5. DiStefano, P. S. and Johnson, E. M. Jr. (1988) Proc. Natl. Acad. Sci. USA 85 (1): 270–274. |
| 6. Vroemen, M. and Weidner, N. (2003) J. Neuroscience Methods 12(2): 135–143. |
| 7. Chalazonitis, A. et al. (1998) Dev. Biol. 204 (2): 385–406. |
| 8. Chambers, S.M. et al. (2009) Nat. Biotechnol. 27 (3): 275–280. |
| 9. Lee, G. et al. (2010) Nat. Protoc. 5 (4): 688–701. |
| 10. Zhou, J. et al. (2010) Stem Cells 28 (10): 1741–1750. |
| 11. Schreiner, C. et al. (2011). Sequential magnetic enrichment of TRA-1-60+ PSCs and CD271+( p75) NCSCs enhances peripheral neuron differentiation. Poster available in the download section. |
| 12. Rozemuller, H. et al. (2010) Stem Cells Dev. 19 (12): 1911–1921. |
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