The authors have no conflicts of interest to disclose.

1. Enrichment of Hematopoietic Progenitor Cells from Adult Whole Blood
NOTE: Hematopoietic progenitor cells or CD34+ cells can be purified from peripheral blood mononuclear cells (PBMCs) derived from a variety of sources including cord blood, leukapheresis material and whole blood using density gradient centrifugation based methods. The method listed here uses whole blood as an example.
<ol>
	<li>Isolation of PBMCs from Whole Blood:
	<ol>
		<li>Add 4.5 ml of lymphocyte separation medium to t...

<ol>
	<li>Azari, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purification+of+immature+neuronal+cells+from+neural+stem+cell+progeny.">Purification of immature neuronal cells from neural stem cell progeny.</a> PLoS ONE. 6, e20941 (2011).</li><li>Azari, H., Sharififar, S., Fortin, J. M., Reynolds, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuroblast+assay:+an+assay+for+the+generation+and+enrichment+of+neuronal+progenitor+cells+from+differentiating+neural+stem+cell+progeny+using+flow+cytometry.">The neuroblast assay: an assay for the generation and enrichment of neuronal progenitor cells from differentiating neural stem cell progeny using flow cytometry.</a> J Vis Exp. (62), (2012).</li><li>Takahashi, K., Yamanaka, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+pluripotent+stem+cells+from+mouse+embryonic+and+adult+fibroblast+cultures+by+defined+factors.">Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors.</a> Cell. 126 (4), 663-676 (2006).</li><li>Takahashi, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+pluripotent+stem+cells+from+adult+human+fibroblasts+by+defined+factors.">Induction of pluripotent stem cells from adult human fibroblasts by defined factors.</a> Cell. 131 (5), 861-872 (2007).</li><li>Zhang, N., An, M. C., Montoro, D., Ellerby, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+Human+Huntington's+Disease+Cell+Model+from+Induced+Pluripotent+Stem+Cells.">Characterization of Human Huntington's Disease Cell Model from Induced Pluripotent Stem Cells.</a> PLoSCurr. 2, RRN1193 (2010).</li><li>Park, I. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disease-Specific+Induced+Pluripotent+Stem+Cells.">Disease-Specific Induced Pluripotent Stem Cells.</a> Cell. 134 (5), 877-886 (2008).</li><li>Song, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+differentiation+of+patient+specific+iPS+cells+as+a+novel+approach+to+study+the+pathophysiology+of+multiple+sclerosis.">Neural differentiation of patient specific iPS cells as a novel approach to study the pathophysiology of multiple sclerosis.</a> Stem Cell Research. 8 (2), 259-273 (2012).</li><li>Vierbuchen, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+conversion+of+fibroblasts+to+functional+neurons+by+defined+factors.">Direct conversion of fibroblasts to functional neurons by defined factors.</a> Nature. 463 (7284), 1035-1041 (2010).</li><li>Pfisterer, U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+conversion+of+human+fibroblasts+to+dopaminergic+neurons.">Direct conversion of human fibroblasts to dopaminergic neurons.</a> Proceedings of the National Academy of Sciences. 108, 10343-10348 (2011).</li><li>Lee, S. -. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Generation+of+Neurosphere-Like+Cells+from+Human+Dermal+Fibroblasts.">Direct Generation of Neurosphere-Like Cells from Human Dermal Fibroblasts.</a> PLoS ONE. 6, e21801 (2011).</li><li>Thier, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Conversion+of+Fibroblasts+into+Stably+Expandable+Neural+Stem+Cells.">Direct Conversion of Fibroblasts into Stably Expandable Neural Stem Cells.</a> Cell Stem Cell. 10 (4), 473-479 (2012).</li><li>Lujan, E., Chanda, S., Ahlenius, H., S&#252;dhof, T. C., Wernig, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+conversion+of+mouse+fibroblasts+to+self-renewing,+tripotent+neural+precursor+cells.">Direct conversion of mouse fibroblasts to self-renewing, tripotent neural precursor cells.</a> Proceedings of the National Academy of Sciences. 109, 2527-2532 (2012).</li><li>Wang, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Derivation+of+neural+stem+cells+from+human+adult+peripheral+CD34++cells+for+an+autologous+model+of+neuroinflammation.">Derivation of neural stem cells from human adult peripheral CD34+ cells for an autologous model of neuroinflammation.</a> PLoS ONE. 8, e81720 (2013).</li><li>Jin, C. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recombinant+Sendai+virus+provides+a+highly+efficient+gene+transfer+into+human+cord+blood-derived+hematopoietic+stem+cells.">Recombinant Sendai virus provides a highly efficient gene transfer into human cord blood-derived hematopoietic stem cells.</a> Gene Ther. 10 (3), 272-277 (2003).</li><li>Ban, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+generation+of+transgene-free+human+induced+pluripotent+stem+cells+(iPSCs)+by+temperature-sensitive+Sendai+virus+vectors.">Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors.</a> Proceedings of the National Academy of Sciences. 108, 14234-14239 (2011).</li><li>Goolsby, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hematopoietic+progenitors+express+neural+genes.">Hematopoietic progenitors express neural genes.</a> Proceedings of the National Academy of Sciences. 100, 14926-14931 (2003).</li></ol>

The study is funded by NIH intramural funds.

A detailed protocol to directly generate neural stem cells from peripheral hematopoietic progenitor cells is provided.
Compared to fibroblast cells, human peripheral blood is more accessible. Using the presented protocol, more than 1 x 105 hematopoietic progenitor cells, or CD34 positive cells, can be enriched from 10 ml of whole blood. Although contamination with platelets is usually not preventable, it can be easily reduced by lower speed centrifugation and it does not interfere w...

In vitro neuronal cultures have been used as a fundamental tool for studies of neurological diseases. Primary animal (mostly rodent) neural cultures1,2 and human neural cell lines derived from gliomas or other tumors are the most commonly used in such studies. However, it has been recognized that there are significant differences between rodent and human cells. Many findings based on rodents cannot be translated to humans. Furthermore, with the rapid developments in analyzing mass genomic information and the relatively easy gene editing and whole genome sequencing, the trend is more and more geared to discovering disease prone genes and delineating their functions and roles in specific diseases, which makes the few human neuronal cell lines have only limited usage. Theoretically, human primary neural cultures derived from samples of patient nervous system are the best choice but they are impossible to obtain; hence alternative methods are necessary. In recent years, some approaches have been pursued, with two being the most distinguishable. Following the development of the technique of generating induced pluripotent stem cells (iPSC) using mouse and human somatic cells3,4, neural cells could be further differentiated from them5-7. However, generating and characterizing iPSC demands intensive labor, techniques, and time input, sometimes even prohibitively. Shortly after, another approach was developed to directly transform neuronal cells from somatic cells8,9. As the resulting neurons are non-proliferative, it limits its application in intensive studies and drug screening, which requires a large amount of cells. To take advantages of both techniques, direct derivation of neural stem/progenitor cells from somatic cells has been explored by several groups10-12, which bypasses the tedious process of iPSC generation and characterization but still provides a decent number of neural stem cells for later neural differentiation. We have previously shown that following the introduction of the Yamanaka transcription factors into hematopoietic progenitor cells, neural stem cells could be directly generated using a neural progenitor cell selecting medium13. Here, we report the method in detail.

<table border="0" cellpadding="0" cellspacing="0" width="584">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Material List</td>
			<td style="width:171px;"></td>
			<td style="width:135px;"></td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Name</td>
			<td style="width:171px;">Company</td>
			<td style="width:135px;">Catalog Number</td>
			<td style="width:72px;">Comments</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Lymphocyte Separation Medium</td>
			<td style="width:171px;">Lonza</td>
			<td style="width:135px;">17-829E</td>
			<td style="width:72px;">1 X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">SepMate</td>
			<td style="width:171px;">Stemcell Technologies</td>
			<td align="right" style="width:135px;">15415</td>
			<td style="width:72px;">15 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Human Serum</td>
			<td style="width:171px;">Invitrogen</td>
			<td style="width:135px;">34005-100</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Antibiotics</td>
			<td style="width:171px;">Gibco</td>
			<td style="width:135px;">15240-062</td>
			<td align="right" style="width:72px;">1%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">CD34 MultiSort Kit</td>
			<td style="width:171px;">Miltenyi Biotec</td>
			<td style="width:135px;">130-056-701</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">EDTA</td>
			<td style="width:171px;">Cellgro</td>
			<td style="width:135px;">46-034-Cl&#160;</td>
			<td style="width:72px;">2mM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">MACS LS Column</td>
			<td style="width:171px;">Miltenyi Biotec</td>
			<td style="width:135px;">130-042-401</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">StemSpan SFEM Medium</td>
			<td style="width:171px;">Stemcell Technologies</td>
			<td align="right" style="width:135px;">9650</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">IMDM</td>
			<td style="width:171px;">Quality Biologicals</td>
			<td style="width:135px;">112-035-101</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">TPO</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">300-18</td>
			<td style="width:72px;">100 ng/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Flt-3</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">300-19</td>
			<td style="width:72px;">100 ng/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">SCF</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">300-07</td>
			<td style="width:72px;">100 ng/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">IL-6</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">200-06</td>
			<td style="width:72px;">20 ng/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">IL-7&#160;</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">200-07</td>
			<td style="width:72px;">20 ng/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">IPS Sendai Reprogramming Kit</td>
			<td style="width:171px;">Life technologies</td>
			<td style="width:135px;">A1378001</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Cell scraper</td>
			<td style="width:171px;">Sarstedt</td>
			<td align="right" style="width:135px;">83.183</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">DMSO</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">D2650</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">CryoTube vials</td>
			<td style="width:171px;">Thermo</td>
			<td align="right" style="width:135px;">368632</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Mr. Frosty container</td>
			<td style="width:171px;">Thermo</td>
			<td style="width:135px;">5100-0001</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">DMEM/F12</td>
			<td style="width:171px;">Life technologies</td>
			<td style="width:135px;">12400-024</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">N2 supplement</td>
			<td style="width:171px;">Life technologies</td>
			<td style="width:135px;">17502-048</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Bovine serum albumin</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">A2934</td>
			<td style="width:72px;">0.1% (w/v)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">bFGF</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">100-18B</td>
			<td style="width:72px;">20 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">EGF</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">AF-100-15</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">B27 supplement</td>
			<td style="width:171px;">Life technologies</td>
			<td style="width:135px;">17504-044</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">NSC serum free medium</td>
			<td style="width:171px;">Life Technologies</td>
			<td style="width:135px;">A1050901</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="40">
			<td height="43" rowspan="2" style="height:43px;width:208px;">Poly-D-lysine/laminin coated cover slips</td>
			<td rowspan="2" style="width:171px;">BD Bioscences</td>
			<td align="right" rowspan="2" style="width:135px;">354087</td>
			<td rowspan="2" style="width:72px;"></td>
		</tr>
		<tr height="3">
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">cAMP</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">A9501</td>
			<td style="width:72px;">300 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Vitamin C</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">A0278</td>
			<td style="width:72px;">0.2 mM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">BDNF</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">450-02</td>
			<td style="width:72px;">10 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">GDNF</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">450-10</td>
			<td style="width:72px;">10 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Poly-L-Ornithine</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">P4957</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">PDGF-AA</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">100-13A</td>
			<td style="width:72px;">10 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">NT-3</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">450-03</td>
			<td style="width:72px;">2 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Shh</td>
			<td style="width:171px;">Peprotech</td>
			<td style="width:135px;">1314-SH/CF</td>
			<td style="width:72px;">2 ng/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">T3</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">T6397</td>
			<td style="width:72px;">3 nM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">PFA</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">P6148</td>
			<td align="right" style="width:72px;">4%</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;width:208px;">PBS</td>
			<td style="width:171px;">Quality Biological</td>
			<td style="width:135px;">119-069-101</td>
			<td style="width:72px;">1X</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">Goat serum</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">G9023</td>
			<td align="right" style="width:72px;">4%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">TritonX-100</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">T9284</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Mouse monoclonal anti-Nestin</td>
			<td style="width:171px;">Millipore</td>
			<td style="width:135px;">AB5922</td>
			<td style="width:72px;">1:1000 dilution</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Anti-SOX2&#160; antibody</td>
			<td style="width:171px;">Applied Stemcell</td>
			<td style="width:135px;">ASA0120</td>
			<td style="width:72px;">Ready to use</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Mouse anti-&#946;III-tubulin antibody</td>
			<td style="width:171px;">Promega</td>
			<td style="width:135px;">G712A</td>
			<td style="width:72px;">1:1000 dilution</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:208px;">Rabbit anti-GFAP antibody</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">G4546</td>
			<td style="width:72px;">1:100 dilution</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Anti-O4 antibody</td>
			<td style="width:171px;">R&#38;D Systems</td>
			<td style="width:135px;">MAB1326</td>
			<td style="width:72px;">IgM; 1 ng/ml</td>
		</tr>
		<tr height="49">
			<td height="49" style="height:49px;width:208px;">Alexa Fluor 594 goat anti-rabbit antibody</td>
			<td style="width:171px;">Life techniologies</td>
			<td style="width:135px;">A11012</td>
			<td style="width:72px;">1:400 dilution</td>
		</tr>
		<tr height="49">
			<td height="49" style="height:49px;width:208px;">Alexa Fluor 488 goat anti-mouse antibody</td>
			<td style="width:171px;">Life techniologies</td>
			<td style="width:135px;">A11001</td>
			<td style="width:72px;">1:400 dilution</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:208px;">Alexa Fluor 488 goat anti-mouse IgM antibody</td>
			<td style="width:171px;">Life techniologies</td>
			<td style="width:135px;">A21042</td>
			<td style="width:72px;">1:250 dilution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:208px;">DAPI</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:135px;">D9542</td>
			<td style="width:72px;">1 ug/ml</td>
		</tr>
	</tbody>
</table>

direct induction of human neural stem cells from peripheral blood hematopoietic progenitor cells

Human disease specific neuronal cultures are essential for generating in vitro models for human neurological diseases. However, the lack of access to primary human adult neural cultures raises unique challenges. Recent developments in induced pluripotent stem cells (iPSC) provides an alternative approach to derive neural cultures from skin fibroblasts through patient specific iPSC, but this process is labor intensive, requires special expertise and large amounts of resources, and can take several months. This prevents the wide application of this technology to the study of neurological diseases. To overcome some of these issues, we have developed a method to derive neural stem cells directly from human adult peripheral blood, bypassing the iPSC derivation process. Hematopoietic progenitor cells enriched from human adult peripheral blood were cultured in vitro and transfected with Sendai virus vectors containing transcriptional factors Sox2, Oct3/4, Klf4, and c-Myc. The transfection results in morphological changes in the cells which are further selected by using human neural progenitor medium containing basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF). The resulting cells are characterized by the expression for neural stem cell markers, such as nestin and SOX2. These neural stem cells could be further differentiated to neurons, astroglia and oligodendrocytes in specified differentiation media. Using easily accessible human peripheral blood samples, this method could be used to derive neural stem cells for further differentiation to neural cells for in vitro modeling of neurological disorders and may advance studies related to the pathogenesis and treatment of those diseases.

The quality of CD34 cells is critical for the success of neural stem cell transduction. High quality CD34 cells proliferate during the first couple of days of culture and appear as non adherent, homogenously round cells, floating just above the bottom of culture vessels (Figure 1A). The successful infection results in cell aggregates, which expands over time (Figure 1B), while non specific cell aggregates may occur during cell culture, that expand and the associations are loose. Adherent...

Watch this Scientific Journal Video about Direct Induction of Human Neural Stem Cells from Peripheral Blood Hematopoietic Progenitor Cells at JoVE.com

Direct Induction of Human Neural Stem Cells from Peripheral Blood Hematopoietic Progenitor Cells

A method was developed to directly derive human neural stem cells from hematopoietic progenitor cells enriched from peripheral blood cells.

Human disease specific neuronal cultures are essential for generating in vitro models for human neurological diseases. However, the lack of access to ...

direct-induction-human-neural-stem-cells-from-peripheral-blood

Research

JoVE Journal

Developmental Biology

12.3K Views.  National Institutes of Health. A method was developed to directly derive human neural stem cells from hematopoietic progenitor cells enriched from peripheral blood cells.