Name: isolation and culture of neural crest stem cells from human hair follicles
Uploaded: 2013-04-06T14:00:00
Description: Watch this Scientific Journal Video about Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles at JoVE.com

这项工作是支持的NIH授予R01AR054593，和R01AR054593-S1徐。

1。组织培养板的制备<ol><li>外套的各孔具有足够的聚-D-赖氨酸（PDL），以覆盖的井的底部。允许板在通风橱中干燥。 </li><li>井干后，用无菌水冲洗，并吸。允许板在通风橱中干燥。 </li><li>当干燥时，纤连蛋白涂层（BioWhittaker公司水溶解于在37℃下过夜，在6毫升的浓度为1毫克）。 </li><li>添加非公务员合约介质[95毫升DMEM/F12，1毫升佩恩/链霉素（P / S），1毫升N2，B272毫升，100μl的巯基乙醇（2ME; 50m...

<ol>
	<li>Cotsarelis, G., Cheng, S. Z., Dong, G., Sun, T. T., Lavker, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Existence+of+slow-cycling+limbal+epithelial+basal+cells+that+can+be+preferentially+stimulated+to+proliferate:+implications+on+epithelial+stem+cells.">Existence of slow-cycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: implications on epithelial stem cells.</a> Cell. 57, 201-209 (1989).</li><li>Cotsarelis, G., Sun, T. T., Lavker, R. 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J. Pathol. 168, 1879-1888 (2006).</li><li>Yu, H., Kumar, S. M., Kossenkov, A. V., Showe, L., Xu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cells+with+neural+crest+characteristics+derived+from+the+bulge+region+of+cultured+human+hair+follicles.">Stem cells with neural crest characteristics derived from the bulge region of cultured human hair follicles.</a> J. Invest. Dermatol. 130, 1227-1236 (2010).</li><li>Wong, C. E., 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+crest-derived+cells+with+stem+cell+features+can+be+traced+back+to+multiple+lineages+in+the+adult+skin.">Neural crest-derived cells with stem cell features can be traced back to multiple lineages in the adult skin.</a> J. Cell Biol. 175, 1005-1015 (2006).</li><li>Sieber-Blum, M., Grim, M., Hu, Y. F., Szeder, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pluripotent+neural+crest+stem+cells+in+the+adult+hair+follicle.">Pluripotent neural crest stem cells in the adult hair follicle.</a> Dev. Dyn. 231, 258-269 (2004).</li><li>Amoh, Y., 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=Implanted+hair+follicle+stem+cells+form+Schwann+cells+that+support+repair+of+severed+peripheral+nerves.">Implanted hair follicle stem cells form Schwann cells that support repair of severed peripheral nerves.</a> Proc. Natl. Acad. Sci. U.S.A. 102, 17734-17738 (2005).</li><li>Koliatsos, V. 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细胞的分离和培养方法的重现性和强大的。我们已经生成的非公务员合约员工从几十个人在广泛的年龄范围。虽然这是最好的，处理组织组织收获后，我们发现头皮组织可以安全地存储在媒体上冰过夜运输对细胞活力的影响最小。 重要的是要与抗生素治疗头皮组织和毛囊分离过程中使用无菌技术，以避免潜在的微生物污染。被丢弃的小改款皮肤或胎儿头皮组织的收益率，数?...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>DMEM</td>
 <td>Invitrogen</td>
 <td>11965-092</td>
 </tr>
 <tr>
 <td>DMEM/F12</td>
 <td>Invitrogen</td>
 <td>11330-32</td>
 </tr>
 <tr>
 <td>Heat-inactivated FBS</td>
 <td>Hyclone</td>
 <td>SH30071.03</td>
 </tr>
 <tr>
 <td>B27 supplement</td>
 <td>Invitrogen</td>
 <td>17504044</td>
 </tr>
 <tr>
 <td>N2 supplement</td>
 <td>Invitrogen</td>
 <td>17502048</td>
 </tr>
 <tr>
 <td>bFGF</td>
 <td>Invitrogen</td>
 <td>PHG0026</td>
 </tr>
 <tr>
 <td>EGF</td>
 <td>R&amp;D system</td>
 <td>236-EG-01M</td>
 </tr>
 <tr>
 <td>IGF-I</td>
 <td>R&amp;D system</td>
 <td>291-G1-050</td>
 </tr>
 <tr>
 <td>0.05% Trypsin/EDTA</td>
 <td>Invitrogen</td>
 <td>25300-054</td>
 </tr>
 <tr>
 <td>Dispase</td>
 <td>Invitrogen</td>
 <td>17105041</td>
 </tr>
 <tr>
 <td>Penicillin-Streptomycin</td>
 <td>Invitrogen</td>
 <td>15070063</td>
 </tr>
 </tbody> 
</table>
Table 1.

isolation and culture of neural crest stem cells from human hair follicles

毛囊进行终身的增长，头发的周期是一个控制的过程，涉及干细胞的增殖和平静。头发隆起是一个很好的成体干细胞的特征利基。这包含了许多不同类型的干细胞，包括上皮干细胞，黑素细胞的干细胞3和神经嵴类似干细胞4-7段的外根鞘。毛囊代表一个可访问的和不同类型的人类干细胞的丰富来源。我们和其他人隔离的神经嵴干细胞（非公务员合约员工）的成人和胎儿毛囊4,5。这些人类干细胞标记的滞留细胞，通过体外细胞不对称分裂，并能够自我更新。他们表示，未成熟的神经嵴细胞的标记物，但不分化标志物。我们的表达谱的研究表明，他们有着相似的基因表达模式与小鼠皮肤未成熟的神经创建ST细胞。它们表现出的克隆的多能在体外克隆单细胞培养后，可以给引起生肌，黑色素细胞和神经元的细胞谱系。分化的细胞，不仅获得了种系特异性标志物，而且在体外条件下表现出适当的功能。此外，这些非公务员合约员工向间充质谱系分化潜能。分化的神经元细胞能坚持在小鼠大脑和保留神经元的分化。它已经表明，毛囊派生的非公务员合约员工可以帮助神经再生，改善运动功能，这些干细胞在小鼠体内移植后，横断性脊髓损伤8。此外，外周神经干细胞移植和植入的皮肤源性坐骨神经神经前体细胞相邻修复，导致髓鞘10。因此，毛囊/皮肤派生的非公务员合约员工已经显示出可喜的结果为r临床前模型egenerative治疗。 体细胞重编程诱导多能干细胞（iPS细胞）已经显示出巨大的潜力，再生医学。不过，也有iPS细胞，特别是基因/病毒整合和潜在的致瘤性多能干细胞的长期影响仍有许多问题没有得到充分解决。还有许多难以克服的障碍前iPS细胞可用于再生医学。鉴于成体干细胞是已知的，是安全的，它们已被用于临床多年，如骨髓移植。很多病人已经从治疗中获益。自体成人干细胞仍然是优选的用于移植细胞。因此，在人的皮肤/头发毛囊容易获得的，可扩展的成体干细胞是再生医学的重要来源。

Watch this Scientific Journal Video about Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles at JoVE.com

Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles

本文提出了一种强大的协议从人的头发毛囊的神经嵴干细胞的分离和培养。

从人类毛囊神经嵴干细胞的分离和培养的

Hair follicles undergo lifelong growth and hair cycle is a well-controlled process involving stem cell proliferation and quiescence. Hair bulge is a ...

Research

JoVE Journal

Neuroscience

Production of Chick Embryo Extract for the Cultivation of Murine Neural Crest Stem Cells

小鼠神经嵴干细胞的培养鸡胚提取物的生产

The neural crest arises from the neuro-ectoderm during embryogenesis and persists only temporarily. Early experiments already proofed pluripotent ...

科研

神经科学

Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells

神经细胞集落形成细胞分析检测，以区分善意的神经祖细胞的神经干细胞

The neurosphere assay (NSA) is one of the most frequently used methods to isolate, expand and also calculate the frequency of neural stem cells (NSCs). ...

Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue

从废弃的人类胎儿大脑皮质组织神经干细胞的生成

Neural stem cells (NSCs) reside along the ventricular zone neuroepithelium during the development of the cortical plate. These early progenitors ...

Isolation and Culture of Human Fungiform Taste Papillae Cells

人力蕈状味觉​​乳头细胞的分离和培养

Taste cells are highly specialized, with unique histological, molecular and physiological characteristics that permit detection of a wide range of simple ...

Isolation and Culture of Rat Embryonic Neural Cells: A Quick Protocol

大鼠胚胎神经细胞的分离和培养：一个快速议定书“

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied ...

Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube

从小鼠胚胎神经管的神经嵴细胞的分离和培养

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to ...

A cGMP-applicable Expansion Method for Aggregates of Human Neural Stem and Progenitor Cells Derived From Pluripotent Stem Cells or Fetal Brain Tissue

人类神经的聚集体cGMP的适用展开法干细胞和祖细胞来源从多能干细胞或胎儿脑组织

A cell expansion technique to amass large numbers of cells from a single specimen for research experiments and clinical trials would greatly benefit the ...

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice

一个鼠标，两种文化：隔离和成人神经的培养干细胞从单个小鼠的两个神经源性区

The neurosphere assay and the adherent monolayer culture system are valuable tools to determine the potential (proliferation or differentiation) of adult ...

Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells

从人多能神经嵴前体细胞的无饲养推导干细胞

Human pluripotent stem cells (hPSCs) have great potential for studying human embryonic development, for modeling human diseases in the dish and as a ...

Isolation and Culture of Adult Neural Stem Cells from the Mouse Subcallosal Zone

分离和成人神经干细胞的培养从鼠标Subcallosal区

Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation ...