The authors wish to thank Harene Ranjithakumaran and Stacey Jackson for technical support. This work was supported in part by grants from the National Health and Medical Research Council (R.C.B. Wong, A. P&#233;bay), the University of Melbourne (R.C.B. Wong), Retina Australia (R.C.B. Wong, S.S.C. Hung, A. P&#233;bay) and the Ophthalmic Research Institute of Australia (R.C.B. Wong, S.S.C. Hung, A. P&#233;bay); Australian Research Council Future Fellowship (A. P&#233;bay, FT140100047), Cranbourne Foundation Fellowship (R.C.B. Wong); intramural funding from the National Institutes for Health (R.C.B. Wong, S.S.C. Hung) and operational infrastructure support from the Victorian Government.

从个人人类头发样品的集合需要通过人的研究伦理委员会在宿主机构伦理委员会批准，并应符合进行与机构准则。 从弹拨头发1.隔离的角质形成细胞<ol><li>解冻细胞外基质（ECM）解决方案（ 即基质胶）在冰O / N。 </li><li>用预冷的枪头，加入200微升的ECM溶液到12毫升冷冻的DMEM / F12培养基。外套一个12孔板用1ml稀释的ECM溶液到每孔中。孵育涂板O / N在37℃。在使用前预热DMEM / F12培养基洗涤包被的平板。 </li><li>通过将手指靠近发根和拔除毛发在一个快速和平滑的运动收集来自个人的头的原代人的头发。检查以确认每个收集的头发中含有一个完整的毛囊并确定联合国的头发的生长阶段明镜解剖显微镜（ 图1）。 <ol><li>收集5-10生长期的毛发从各个提取角质。过程弹拨毛发尽快确保角化细胞的成功的推导。 注意： 图1示出在生长周期的不同阶段的毛发的形态。毛发生长初期是在生长阶段，上皮周围的头发的长度，称为ORS的可见外层。另一方面，休止期头发代表头发在休止期和不具有可见的ORS，但有一个明确的细胞覆盖的毛发的根，被称为休止凸起的球。 </li></ol></li><li>将头发样品到含有10ml抗生素混合料在室温洗5分钟的培养皿。 </li><li>用消毒剪刀和镊子，剪除过多的毛干，留下了头发的片段用一个完整的毛囊和周围0.5 - 毛干为1厘米。 </li><li>仔细TRAnsfer 1发片段插入使用镊子将ECM涂覆的板的各孔中。 </li><li>使用1毫升吸管，加入约3滴敲除血清替代品（KSR）培养基（〜100 - 200微升）到头发样品。 注意：保持头发的片段密切接触板面，以允许附件和随后的角质细胞生长。加入过量的介质，在该步骤中可以使头发片段浮起并减小成功头发附着的机会。 </li><li>孵育头发样品在37℃的培养箱中5％的CO 2，并允许毛囊附着O / N。第二天，轻轻再添3滴KSR介质（100〜 - 200微升），以保持头发湿润的样品。 </li><li>观察该头发样本每日以确认毛囊已经成功附着。通常附着毛囊内1-3天明显。厚厚的毛发通常更容易附着相比细毛，因为细毛有较高倾向浮起阻碍连接过程。 <ol><li>一旦头发已经附着到涂板，加入500μl的KSR培养基到孔中。添加媒体时头发样品可能没有牢固地附着要格外小心。从这一点来说，改变媒体每2天。 注：后3-7天（ 图2A-B）副产物角质应该可以看到从发片段的ORS区域。让角质形成细胞在下一节介绍的传代细胞前长到14天。 </li></ol></li></ol> 2.维护和角质形成细胞传代<ol><li>预涂层用涂料矩阵包一个6孔板。添加680微升稀释介质的从所述试剂盒向每个孔，接着6.8微升涂布基质溶液。摇板，以确保在平板上涂料基液的均匀分布。 </li><li>允许涂覆的板孵育30分钟，在室温。拆除之前的角质形成细胞的电镀镀层Matrix解决方案。 </li><li>离解的角质形成细胞为单细胞悬浮液，用500μL每孔0.25％胰蛋白酶的为2-5分钟，在37℃。灭活用500μl含培养基的血清（ 例如，胎牛血清（FBS）的培养基）的胰蛋白酶，并收集到无菌的15毫升管。删除使用消毒镊子头发的片段。 </li><li>确定使用血球收获的细胞的数目。 </li><li>在200离心细胞3分钟×g下。洗涤细胞，用PBS和重复离心。吸出PBS，重悬细胞沉淀在新鲜的角质形成细胞培养基。 </li><li>板向下4×10 4的角质形成细胞进入ECM涂覆后的板在角质形成细胞培养基的存在下的一个阱。保持角化细胞在37℃培养箱中，用5％的CO 2（ 图2C）。改变媒体每天都和传代细胞时，文化是〜80％汇合，WHI通道可能需要约2-7天，取决于角化细胞的传代次数和细胞增殖率。派生角质应该扩大使用标准慢速冷冻方法冷冻保存16产生足够的冷冻库存。 注意：所述的培养条件下支持角化细胞具有典型的鹅卵石形态（ 图2C）的同源性的群体的膨胀。偶尔，一些差异化的角质形成细胞，具有大而扁平的形态也失去复制能力，可以在培养中观察到。角化细胞的表征可以通过免疫染色采用抗细胞角蛋白14来执行，如前所述17。支原体后使用市售的试剂盒1-3通道建立角化细胞系也应进行测试。 </li></ol> 3.从一代人iPS细胞角质形成细胞利用游离型载体<ol><li>预涂层的6孔板0.1％明胶（2毫升/孔）。允许明胶涂覆至少20分钟，在室温。 </li><li>对于重编程的实验中，使用角质形成不迟于通道6离解角质成单细胞悬浮液与1ml每孔0.25％胰蛋白酶为2-5分钟，在37℃。灭活胰蛋白酶用1毫升含有培养基的血清（ 例如，胎牛血清介质）的，并收集在一个15毫升管。 </li><li>离心细胞3分钟，在200×g离心和重悬细胞的角质形成细胞培养基中。每孔板1×10 5 -1.5×10 5角朊细胞的糊化板O / N。 </li><li>于次日（第0天），检查板，以保证细胞50-60％汇合。 注意：如果角质小于〜50％汇合，他们转染后也可能不会增长。 </li><li>在第0天，执行对角化细胞的游离型重编程载体的转染，使用8微升转染试剂的比率为2微克总质粒DNA。<ol><li>以0.5微克的每个质粒4（pCXLE-EGFP，pCXLE-hOct3 / 4-shP53F，pCXLE-HSK，pCXLE-讫）和稀释的100微升减少血清培养基（ 如，OPTI-MEM）的。 注意：质粒pCXLE-eGFP的用于监视在角化细胞的转染效率和不需要成功重新编程。 </li><li>添加8微升转染试剂来稀释的DNA。轻弹管混合。允许该DNA反应混合物休息原状在室温15分钟。 </li><li>更换角化细胞培养基用新鲜培养基。 </li><li>轻轻的DNA反应混合物用5％的CO 2添加到角质细胞培养和孵化在37℃培养箱4小时。 注意：不建议在执行角质形成细胞转染延长O / N，因为他们表现出低生存染后。 </li><li> 4小时后转染后，更换角化细胞培养基用新鲜培养基。 </li></ol></li><li>在第1天（第1天后反染），改变角化细胞培养基中并通过估计在荧光显微镜下的eGFP阳性细胞百分比检查转染效率。通常&gt; 60％-70％的转染效率观察使用该协议的主要角质形成细胞。 </li><li>第2天，按步骤3.5节重复转染。连续两次转染通常会达到&gt; 90％的转染效率。角质形成细胞培养通常是由第二次转染的端部到达汇合。 <ol><li>制备小鼠胚胎成纤维细胞（MEF）的送料器的10cm皿的每个重编程反应。准备有丝分裂失活的MEF饲养如前所述18。 </li><li>预涂层10cm皿用5ml 0.1％明胶的至少20分钟。板4×10 6％的FBS媒体10 cm培养皿有丝分裂灭活的MEF。使细胞沉降并附着O / N在37℃培养箱，用5％的CO 2。 </li></ol></li><li>在第3天，哈尔如在步骤3.2和3.3中所述归属用0.25％胰蛋白酶的角质形成细胞。重悬在KSR培养基中的角质形成细胞和板90％，从1以及一个6孔板的角质形成细胞的成10cm的MEF饲养皿KSR培养基。 </li><li>从第4天起，每天换了KSR媒介。 <ol><li>或者18天之后，更换培养基每隔一天。 注：非重编程角化细胞将停止增殖，同时具有限定边界，类似于人类胚胎干细胞（胚胎干细胞）hiPSC菌落会从第14天 - 21。 </li></ol></li><li>从21日起隔离hiPSC殖民地人工清扫。避免hiPSC殖民地，它们紧密地聚集在一起，只能挑hiPSC殖民地有明确从别人分开。 注：hiPSC殖民地可以采摘较早，但与定义的边界殖民地可能不是很明显，在较早的时间点给小菌落大小。重编程效率差异可能会有所不同erent病人的角质形成细胞，并且还通过其他因素如通道数或角质形成细胞的增殖率的影响。 <ol><li>对于hiPSC殖民地人工清扫，使用26G针头切解剖显微镜下hiPSC殖民地。切hiPSC殖民地到约300小块 - 600微米长。转让从一个殖民地hiPSC件到一个新的MEF饲养板（2.5×10 4 MEF /厘米2）建立iPS细胞的克隆系。最初，文化的12孔板或器官培养的菜肴之一以及每个hiPSC克隆行，然后扩展到6孔板格式。 </li><li>维持iPS细胞在KSR介质上MEF饲养建立无性系。一旦培养达到70％ - 80％的汇合与hiPSC菌落〜1.5mm的直径，使用根据制造商的说明标准的酶传代方法（的Accutase，分散酶或胶原酶IV）的通道人iPS细胞。 </li></ol></li><li>建立特征线hiPSCs到确认多能性及其分化成细胞代表在体外和/或体内 13,19三种胚层的能力。此外，使用市售的试剂盒定期测试建立hiPSC线支原体，以确保它们是无病原体。定期监测的hiPSC线通过核型分析和基于阵列的拷贝数变异（CNV）分析基因组稳定性。 </li></ol>

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+is+the+point+of+large-scale+collections+of+human+induced+pluripotent+stem+cells.">What is the point of large-scale collections of human induced pluripotent stem cells.</a> Nat Biotechnol. 31, 875-877 (2013).</li><li>Utikal, 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=Immortalization+eliminates+a+roadblock+during+cellular+reprogramming+into+iPS+cells.">Immortalization eliminates a roadblock during cellular reprogramming into iPS cells.</a> Nature. 460, U1145-U1112 (2009).</li><li>Xu, 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=Proliferation+rate+of+somatic+cells+affects+reprogramming+efficiency.">Proliferation rate of somatic cells affects reprogramming efficiency.</a> J Biol Chem. 288, 9767-9778 (2013).</li><li>Liu, 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=Late+passage+human+fibroblasts+induced+to+pluripotency+are+capable+of+directed+neuronal+differentiation.">Late passage human fibroblasts induced to pluripotency are capable of directed neuronal differentiation.</a> Cell Transplant. 20, 193-203 (2011).</li><li>Huallachain, M., Karczewski, K. J., Weissman, S. M., Urban, A. E., Snyder, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extensive+genetic+variation+in+somatic+human+tissues.">Extensive genetic variation in somatic human tissues.</a> Proc Natl Acad Sci U S A. 109, 18018-18023 (2012).</li><li>Abyzov, A., 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=Somatic+copy+number+mosaicism+in+human+skin+revealed+by+induced+pluripotent+stem+cells.">Somatic copy number mosaicism in human skin revealed by induced pluripotent stem cells.</a> Nature. 492, 438-442 (2012).</li></ol>

The authors have no conflict of interest.

患者特异性人iPS细胞的产生提供了用于在体外研究发病在患病细胞类型的独特方法，并且还提供了用于药物筛选平台，以确定新的分子，可以拯救疾病表型。使用人iPS细胞这种疾病建模方法已取得了令人鼓舞的结果，适用于各种疾病，包括长QT综合征，亨廷顿氏病，帕金森氏病和肌萎缩性侧索硬化22。若干举措已经展开，建立以病人特异性iPS细胞，其中包括在美国，欧洲，澳大利亚?...

人类诱导多能干细胞（人iPS细胞）的发现已经彻底改变了再生医学的领域中，提供用于产生患者特异性干细胞1-3的一种可行的方法。人iPS细胞已经成功地从不同的体细胞类型，包括成纤维细胞4,5，造血细胞6,7，从尿8肾小管上皮细胞和角质形成细胞9,10产生的。迄今为止，皮肤成纤维细胞和造血细胞代表了最常用的细胞来源，用于产生病人的具体的iPSC。可以说，这是由于这样的事实，即皮肤活检和采血是常规的医疗程序，并已在许多国家建立大型生物库患者血液或皮肤的样品。 相较于血细胞和皮肤成纤维需要侵入性的提取方法，角质代表一个方便的细胞类型的hiPSC产生。 KeratinocytES是形成皮肤的外表皮屏障，并且还发现，在指甲和头发角蛋白11的富上皮细胞。特别是，角朊细胞可以在毛囊，即覆盖毛干连同内根鞘（IRS）的细胞（12， 图1）的外部的蜂窝层的外根鞘（ORS）中找到。由于毛发收集是一个简单的过程，不需要医务人员的协助下，它提供了一个机会，让患者收集并发送自己的头发样品的实验室，这将极大地方便了hiPSC代病人样本的采集。表皮角化细胞也有较高的重编程效率和更快的重编程动力学相比的成纤维细胞，加入到用角质形成细胞作为起始细胞hiPSC代9,13的优点。此外，人iPS细胞还可以使用毛囊内其他细胞群产生，包括位于毛囊14,15的底部的毛乳头细胞。 IPSC的代发使用来源的细胞以前的报告中经常使用逆转录病毒或慢病毒为基础的重编程方法9,14,15。然而，这些病毒方法的重新编程过程中引进国外转基因不良基因组整合。相比较而言，使用游离型载体的代表一个可行的，非病毒重新编程方法来生成积分-自由iPSCs的4。我们以前曾开发出一种简单，经济有效和非病毒的方法来有效地重新编程角化细胞到使用附加型载体13人iPS细胞。在这里，我们提供的角质细胞衍生的人iPS细胞，包括来自弹拨头发，扩展和维护的角质形成和随后的重新编程，以产生iPS细胞的角质形成细胞的推导的生成的详细协议。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Antibiotic Mix:&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>250 ng/ml Antimycotic amphotericin B</td>
			<td>Sigma</td>
			<td>A2942-20ml</td>
			<td>Antibiotic mix is made up in PBS.&#160;</td>
		</tr>
		<tr>
			<td>1X Penicillin/Streptomycin</td>
			<td>Invitrogen</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS (-)</td>
			<td>Invitrogen</td>
			<td>14190-144</td>
			<td></td>
		</tr>
		<tr>
			<td>Knockout Serum Replacement (KSR) medium:&#160;</td>
			<td></td>
			<td></td>
			<td>KSR medium is filtered using Stericup (Millipore, #SCGPU05RE) before use. bFGF is added fresh to the media before use.</td>
		</tr>
		<tr>
			<td>20% knockout serum replacement (KSR)</td>
			<td>Invitrogen</td>
			<td>10828-028</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F12 with glutamax</td>
			<td>Invitrogen</td>
			<td>10565-042</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#215; MEM non-essential amino acid</td>
			<td>Invitrogen</td>
			<td>11140-050</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;0.5&#215; Penicillin/Streptomycin</td>
			<td>Invitrogen</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;0.1 mM &#946;-mercaptoethanol</td>
			<td>Invitrogen</td>
			<td>21985</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;bFGF (10 ng/ml, added fresh)</td>
			<td>Millipore</td>
			<td>GF003</td>
			<td></td>
		</tr>
		<tr>
			<td>Keratinocyte medium:&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;EpiLife with 60&#160;&#181;M Calcium</td>
			<td>Invitrogen</td>
			<td>M-EPI-500-CA</td>
			<td></td>
		</tr>
		<tr>
			<td>1&#215; Human keratinocyte growth supplement (HKGS)</td>
			<td>Invitrogen</td>
			<td>S-001-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS) medium:&#160;</td>
			<td></td>
			<td></td>
			<td>FBS medium is filtered using Stericup (Millipore, #SCGPU05RE)&#160; before use.</td>
		</tr>
		<tr>
			<td>10% fetal bovine serum (FBS)</td>
			<td>Invitrogen</td>
			<td>26140079</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM&#160;</td>
			<td>Invitrogen</td>
			<td>11995-073</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5&#215; Penicillin/Streptomycin</td>
			<td>Invitrogen</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>2 mM L-glutamine</td>
			<td>Invitrogen</td>
			<td>25030</td>
			<td></td>
		</tr>
		<tr>
			<td>0.25% trypsin-EDTA</td>
			<td>Invitrogen</td>
			<td>25200-056</td>
			<td></td>
		</tr>
		<tr>
			<td>Extracellular Matrix (ECM):</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Matrigel</td>
			<td>Corning&#160;</td>
			<td>354234</td>
			<td>Aliquot Matrigel stock and store in -80&#176;C following manufacturer&#8217;s instructions. Stock concentration of Matrigel varies slightly from batch to batch (~9mg/ml). We recommend to use 200&#181;l matrigel for coating a 12-well plate (~150&#181;g/well).&#160;</td>
		</tr>
		<tr>
			<td>Coating Matrix Kit&#160;</td>
			<td>Invitrogen</td>
			<td>R-011-K</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasmids:&#160;</td>
			<td></td>
			<td></td>
			<td rowspan="5">Note that pCXLE-eGFP is only used for monitoring transfection efficiency and is not required for reprogramming.</td>
		</tr>
		<tr>
			<td>-&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; pCXLE-eGFP</td>
			<td>Addgene</td>
			<td>27082</td>
		</tr>
		<tr>
			<td>-&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; pCXLE-hOct3/4-shP53F</td>
			<td>Addgene</td>
			<td>27077</td>
		</tr>
		<tr>
			<td>-&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; pCXLE-hSK</td>
			<td>Addgene</td>
			<td>27078</td>
		</tr>
		<tr>
			<td>-&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; pCXLE-hUL</td>
			<td>Addgene</td>
			<td>27080</td>
		</tr>
		<tr>
			<td>Transfection reagent Fugene HD</td>
			<td>Promega</td>
			<td>E231B</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin (from porcine skin)</td>
			<td>Sigma</td>
			<td>G1890</td>
			<td>Make up 0.1% gelatin in distilled water. Autoclave before use.&#160;</td>
		</tr>
		<tr>
			<td>Reduced Serum medium: OPTI-MEM</td>
			<td>Invitrogen</td>
			<td>31985062</td>
			<td></td>
		</tr>
		<tr>
			<td>Accutase</td>
			<td>Sigma</td>
			<td>A6964-100ml</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse embryonic fibroblast (MEF) feeder</td>
			<td></td>
			<td></td>
			<td>MEF can be inactivated by mitomycin C treatment or irradiation as described previously 16.</td>
		</tr>
		<tr>
			<td>26G needle</td>
			<td>Terumo</td>
			<td>NN2613R</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well plate (tissue culture treated)</td>
			<td>BD Biosciences</td>
			<td>353046</td>
			<td></td>
		</tr>
		<tr>
			<td>12-well plate (tissue culture treated)</td>
			<td>BD Biosciences</td>
			<td>353043</td>
			<td></td>
		</tr>
		<tr>
			<td>10 cm dish (tissue culture treated)</td>
			<td>BD Biosciences</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr>
			<td>Dispase</td>
			<td>Invitrogen</td>
			<td>17105-041</td>
			<td>Use at 10mg/ml</td>
		</tr>
		<tr>
			<td>Collagenase IV</td>
			<td>Invitrogen</td>
			<td>17104-019</td>
			<td>Use at 1mg/ml</td>
		</tr>
		<tr>
			<td>TRA-160 antibody</td>
			<td>Millipore</td>
			<td>MAB4360</td>
			<td>Use at 5&#181;g/ml</td>
		</tr>
		<tr>
			<td>OCT4 antibody</td>
			<td>Santa Cruz</td>
			<td>SC-5279</td>
			<td>Use at 5&#181;g/ml</td>
		</tr>
		<tr>
			<td>NANOG antibody</td>
			<td>R&#38;D Systems</td>
			<td>AF1997</td>
			<td>Use at 10&#181;g/ml</td>
		</tr>
		<tr>
			<td>MycoAlert Detection kit</td>
			<td>Lonza</td>
			<td>LT07-418</td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of integration-free human induced pluripotent stem cells using hair-derived keratinocytes

Recent advances in reprogramming allow us to turn somatic cells into human induced pluripotent stem cells (hiPSCs). Disease modeling using patient-specific hiPSCs allows the study of the underlying mechanism for pathogenesis, also providing a platform for the development of in vitro drug screening and gene therapy to improve treatment options. The promising potential of hiPSCs for regenerative medicine is also evident from the increasing number of publications (>7000) on iPSCs in recent years. Various cell types from distinct lineages have been successfully used for hiPSC generation, including skin fibroblasts, hematopoietic cells and epidermal keratinocytes. While skin biopsies and blood collection are routinely performed in many labs as a source of somatic cells for the generation of hiPSCs, the collection and subsequent derivation of hair keratinocytes are less commonly used. Hair-derived keratinocytes represent a non-invasive approach to obtain cell samples from patients. Here we outline a simple non-invasive method for the derivation of keratinocytes from plucked hair. We also provide instructions for maintenance of keratinocytes and subsequent reprogramming to generate integration-free hiPSC using episomal vectors.

毛发经过生长周期3个不同阶段：生长期（生长阶段），退行期（回归阶段）和休止期（休息阶段）20,21。生长期毛囊中含有上皮多层;这些层包括ORS，IRS和毛干（ 图1）。毛发生长初期最终经历过渡到退行期阶段，该阶段的特点是ORS的和终止的毛干分化的细胞凋亡。最后，退行期的头发过渡到休止期，在细胞凋亡停止，休止期毛囊变得与静态特征休止期隆起20,21。 <p ...

Watch this Scientific Journal Video about Generation of Integration-free Human Induced Pluripotent Stem Cells Using Hair-derived Keratinocytes at JoVE.com

Generation of Integration-free Human Induced Pluripotent Stem Cells Using Hair-derived Keratinocytes

This manuscript provides a step-by-step procedure for the derivation and maintenance of human keratinocytes from plucked hair and subsequent generation of integration-free human induced pluripotent stem cells (hiPSCs) by episomal vectors.

集成的无人类诱导多能干细胞的生成方法头发衍生角质形成细胞

Recent advances in reprogramming allow us to turn somatic cells into human induced pluripotent stem cells (hiPSCs). Disease modeling using ...

generation-integration-free-human-induced-pluripotent-stem-cells

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Developmental Biology

11.4K 次观看.  University of Melbourne. This manuscript provides a step-by-step procedure for the derivation and maintenance of human keratinocytes from plucked hair and subsequent generation of integration-free human induced pluripotent stem cells (hiPSCs) by episomal vectors. 