Name: isolation, culture, and characterization of primary schwann cells, keratinocytes, and fibroblasts from human foreskin
Uploaded: 2022-03-23T15:00:00
Description: Watch this Scientific Journal Video about Isolation, Culture, and Characterization of Primary Schwann Cells, Keratinocytes, and Fibroblasts from Human Foreskin at JoVE.com

我们要感谢Fadia Kamal博士和Reyad Elbarbary博士允许我们使用实验室仪器和技术支持。这项工作得到了NIH（K08 AR060164-01A）和国防部（W81XWH-16-1-0725）向J.C.E.的资助，以及宾夕法尼亚州立大学好时医学中心的机构支持。

出于研究目的的去识别化人类包皮组织的获取和使用进行了审查，并获得了宾夕法尼亚州立大学医学院机构审查委员会（IRB #17574）确定的“非人类研究”。
注意：通过遵循以下方案，从单个包皮中获得2.4 x 106 角质形成细胞，4.4 x 106 成纤维细胞和1.1 x 106 SC。通常，这些原代细胞可用于3次传代，具体取决于实验条件。
1. 原代?...

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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=Advantages+and+disadvantages+of+using+human+cells+for+pharmacological+and+toxicological+studies.">Advantages and disadvantages of using human cells for pharmacological and toxicological studies.</a> Human &amp; Experimental Toxicology. 13 (8), 568-573 (1994).</li><li>Green, H., Kehinde, O., Thomas, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+of+cultured+human+epidermal+cells+into+multiple+epithelia+suitable+for+grafting.">Growth of cultured human epidermal cells into multiple epithelia suitable for grafting.</a> Proceedings of the National Academy of Sciences of the United States of America. 76 (11), 5665-5668 (1979).</li><li>Fuchs, E., Green, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+keratin+gene+expression+during+terminal+differentiation+of+the+keratinocyte.">Changes in keratin gene expression during terminal differentiation of the keratinocyte.</a> Cell. 19 (4), 1033-1042 (1980).</li><li>Aasen, T., Izpisua Belmonte, J. 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R., Cheret, J., Yosipovitch, G., Paus, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Schwann+cells+as+underestimated,+major+players+in+human+skin+physiology+and+pathology.">Schwann cells as underestimated, major players in human skin physiology and pathology.</a> Experimental Dermatology. 29 (1), 93-101 (2020).</li><li>Laverdet, 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=Skin+innervation:+important+roles+during+normal+and+pathological+cutaneous+repair.">Skin innervation: important roles during normal and pathological cutaneous repair.</a> Histology and Histopathology. 30 (8), 875-892 (2015).</li><li>Jessen, K. R., Mirsky, R., Lloyd, A. 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该协议描述了一种从包皮的单个片段中分离三个不同细胞群的方法，即角质形成细胞，成纤维细胞和施旺细胞。有一些分离方案可用于分离角质形成细胞和成纤维细胞2，3，5，6，但没有一种描述SC分离。除了皮肤中的关键结构细胞、角质形成细胞和成纤维细胞外，皮肤还受到感觉传入结构和自主神经?...

衍生自活组织并在 体外 条件下培养的原代细胞与生理状态1非常相似，使其成为研究生理和病理生理过程的理想模型。皮肤含有多种细胞类型，包括角质形成细胞，成纤维细胞，皮脂细胞，黑素细胞和施万细胞（SC），可以分离和培养用于 体外 实验。尚未描述从单块皮肤中分离和培养角质形成细胞、成纤维细胞和 SC 的方法。该协议的目标是双重的：1）建立一种可靠且可重复的真皮SC分离和培养的方法，以及2）使用一种有效，稳健的方法从单个人包皮中分离角质形成细胞，成纤维细胞和SC。
目前，已有分离皮肤角质形成细胞2，3，4和成纤维细胞5，6的既定方案。这些研究描述了从皮肤中分离角质形成细胞，成纤维细胞或两者，但没有方案解决如何从人类皮肤建立原代SC的培养物。最近的研究表明，神经元SCs调节角质形成细胞和成纤维细胞过程并调节正常的皮肤生理功能7。因此，SC对皮肤稳态至关重要，并且对影响相邻皮肤细胞类型行为的调节生理学有重大贡献8。因此，允许分离每种细胞类型的方案非常适合涉及细胞 - 细胞间通信或细胞类型之间串扰的体外实验。
该协议描述了从单个皮肤建立原代细胞的单个细胞培养物。当可用组织量有限时，该方案特别有用。此外，从单个供体中分离所有三种细胞类型可以在细胞类型或共培养实验之间进行强有力的比较，同时减轻所需实验期间遗传学的影响。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#181;M sterile filters (Millex-GP Syringe Filter Unit,polyethersulfone)</td>
			<td>MilliporeSigma</td>
			<td>SLGPR33RS</td>
			<td></td>
		</tr>
		<tr>
			<td>70 &#181;M cell strainers</td>
			<td>CELLTREAT</td>
			<td>229483</td>
			<td></td>
		</tr>
		<tr>
			<td>100 &#181;M cell strainers</td>
			<td>CELLTREAT</td>
			<td>229485</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL disposable syringes</td>
			<td>BD Luer-Lok</td>
			<td>BD-309659</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL disposable syringes (Syringe sterile, single use)</td>
			<td>BD Luer-Lok</td>
			<td>BD309646</td>
			<td></td>
		</tr>
		<tr>
			<td>10 mL disposable syringes</td>
			<td>BD Luer-Lok</td>
			<td>BD305462</td>
			<td></td>
		</tr>
		<tr>
			<td>1% TritonX-100</td>
			<td>Sigma</td>
			<td>X100-1L</td>
			<td>Prepared at the time of use</td>
		</tr>
		<tr>
			<td>4% paraformaldehyde solution</td>
			<td>ThermoFisher Scientific</td>
			<td>J19943.K2</td>
			<td>Ready to use and store at 4 &#176;C</td>
		</tr>
		<tr>
			<td>5% BSA</td>
			<td>Sigma</td>
			<td>A7906-100G</td>
			<td>Prepared at the time of use</td>
		</tr>
		<tr>
			<td>70% ethanol</td>
			<td>Pharmco</td>
			<td>111000200</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic</td>
			<td>ScienCell Research</td>
			<td>503</td>
			<td></td>
		</tr>
		<tr>
			<td>Chemometec Vial1-Cassette</td>
			<td>Fisher Scientific</td>
			<td>NC1420193</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagenase</td>
			<td>Gibco</td>
			<td>17018-029</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Fisherbrand</td>
			<td>12544D 22*50-1.5</td>
			<td></td>
		</tr>
		<tr>
			<td>Dispase I</td>
			<td>Sigma-Aldrich</td>
			<td>D46693</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM basal medium</td>
			<td>ScienCell Research</td>
			<td>9221</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s phosphate-buffered saline free from Ca2+ and Mg2+ (DPBS)</td>
			<td>Corning</td>
			<td>&#160;21-031-CV)</td>
			<td></td>
		</tr>
		<tr>
			<td>Nunc 15 mL Conical Sterile Polypropylene Centrifuge Tubes</td>
			<td>ThermoFisher Scientific</td>
			<td>339651</td>
			<td></td>
		</tr>
		<tr>
			<td>Nunc 50 mL Conical Sterile Polypropylene Centrifuge Tubes</td>
			<td>ThermoFisher Scientific</td>
			<td>339653</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 mL micro-centrifuge tubes</td>
			<td>Fisherbrand</td>
			<td>02-681-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum (FBS)</td>
			<td>ThermoFisher Scientific</td>
			<td>10082147</td>
			<td></td>
		</tr>
		<tr>
			<td>Fibroblast complete medium</td>
			<td>ScienCell Research</td>
			<td>2331</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594</td>
			<td>Invitrogen</td>
			<td>A11032</td>
			<td>Dilution (1:500)</td>
		</tr>
		<tr>
			<td>Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td>Dilution (1:500)</td>
		</tr>
		<tr>
			<td>Hanks&#39; Buffered Saline Solution (HBSS buffer)</td>
			<td>Lonza</td>
			<td>CC-5022</td>
			<td></td>
		</tr>
		<tr>
			<td>Human foreskin</td>
			<td></td>
			<td></td>
			<td>De-identified human foreskin tissue for research purposes (Institutional Review Board- IRB #17574).</td>
		</tr>
		<tr>
			<td>KGM-GOLD keratinocyte medium (KGM gold and supplements)</td>
			<td>Lonza</td>
			<td>00192151 and 00192152</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse alpha-smooth muscle actin antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>14-9760-82</td>
			<td>Dilution (1:200)</td>
		</tr>
		<tr>
			<td>Mouse Cytokeratin14 antibody</td>
			<td>Abcam</td>
			<td>ab7800</td>
			<td>Dilution (1:100)</td>
		</tr>
		<tr>
			<td>Mouse S100 antibody</td>
			<td>ThermoFisher Scientific</td>
			<td>MA5-12969</td>
			<td>Dilution (1:200)</td>
		</tr>
		<tr>
			<td>Multi chambered (4 well glass slide)</td>
			<td>Tab-Tek</td>
			<td>154526</td>
			<td></td>
		</tr>
		<tr>
			<td>NucleoCounter -Via1-Cassette</td>
			<td>Chemometec</td>
			<td>941-0012</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly-L-Lysisne (PLL)</td>
			<td>ScienCell Research</td>
			<td>32503</td>
			<td></td>
		</tr>
		<tr>
			<td>ProLong Gold Anti-fade Mountant with DAPI</td>
			<td>Invitrogen</td>
			<td>P36935</td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit K10 antibody</td>
			<td>Sigma-Aldrich</td>
			<td>SAB4501656</td>
			<td>Dilution (1:100)</td>
		</tr>
		<tr>
			<td>Rabbit p75-NTR antibody</td>
			<td>Millipore</td>
			<td>AB1554</td>
			<td>Dilution (1:500)</td>
		</tr>
		<tr>
			<td>Rabbit vimentin</td>
			<td>ProteinTech</td>
			<td>10366-1-AP</td>
			<td>Dilution (1:200)</td>
		</tr>
		<tr>
			<td>Schwann cell culture medium</td>
			<td>ScienCell Research</td>
			<td>1701</td>
			<td></td>
		</tr>
		<tr>
			<td>Precision tweezers DUMONT straight with extra fine tips Dumostar, 5</td>
			<td>ROTH</td>
			<td>LH75.1</td>
			<td>Sterilize with 70% alcohol before use</td>
		</tr>
		<tr>
			<td>IRIS Scissors, sharp/sharp. Length 4&#8211;3/8&#34;(111mm)</td>
			<td>Codman</td>
			<td>54-6500</td>
			<td>Sterilize with 70% alcohol before use</td>
		</tr>
		<tr>
			<td>Sterilized surgical - sharp blade (Duro Edge Economy Single Edge Blades)</td>
			<td>Razor blade company</td>
			<td>94-0120</td>
			<td>Sterilize with 70% alcohol before use</td>
		</tr>
		<tr>
			<td>T25 culture flask</td>
			<td>Corning</td>
			<td>353109</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin neutralization buffer (TNS)</td>
			<td>Lonza</td>
			<td>CC-5002</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin/EDTA</td>
			<td>Lonza</td>
			<td>CC-5012</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>ZEISS</td>
			<td>Axio Observer 7- Axiocam 506 mono &#8211; Apotome.2 microscope</td>
			<td>For immunofluorescence of chamber slide containing stained cells</td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>ZEISS</td>
			<td>Primovert</td>
			<td>For visulaizing/observing cell attachment or detachment</td>
		</tr>
	</tbody>
</table>

isolation, culture, and characterization of primary schwann cells, keratinocytes, and fibroblasts from human foreskin

该协议描述了分离方法，培养条件以及使用皮肤的快速酶解离具有高产量和活力的人类原代细胞的表征。原代角质形成细胞、成纤维细胞和施万细胞都是从人类新生儿包皮中采集的，可按照标准护理程序获得。去除的皮肤被消毒，皮下脂肪和肌肉被用手术刀去除。该方法包括表皮和真皮层的酶促和机械分离，然后进行额外的酶消化，以从这些皮肤层中的每一层获得单细胞悬浮液。最后，按照标准的细胞培养方案在适当的细胞培养基中培养单细胞，以在几周内保持生长和活力。总之，这种简单的方案允许从一块皮肤中分离，培养和表征所有三种细胞类型，以进行皮肤 - 神经模型的 体外 评估。此外，这些细胞可以在共培养物中一起使用，以测量它们对彼此的影响以及它们对 体外 创伤的反应，其形式是在与伤口愈合相关的培养物中机器人进行的划痕。

正常新生儿包皮用于原发性表皮角质形成细胞和真皮SC和成纤维细胞的分离。将分离的原代细胞在含有生长因子的相应细胞培养基中培养。在培养瓶中接种SCs和成纤维细胞后，大多数细胞在2小时内粘附在烧瓶底部。在角质形成细胞的情况下，大多数角质形成细胞粘附24小时。分离的表皮原代角质形成细胞在第7天达到85%汇合，并表现出特征性细胞形态（鹅卵石形状）（图1A）?...

Watch this Scientific Journal Video about Isolation, Culture, and Characterization of Primary Schwann Cells, Keratinocytes, and Fibroblasts from Human Foreskin at JoVE.com

Isolation, Culture, and Characterization of Primary Schwann Cells, Keratinocytes, and Fibroblasts from Human Foreskin

研究与肌肉骨骼损伤相关的伤口愈合通常需要评估施万细胞（SCs）、角质形成细胞和成纤维细胞之间的 体外 相互作用。该协议描述了从人包皮中分离，培养和表征这些原代细胞。

从人包皮中分离、培养和表征原代施旺细胞、角质形成细胞和成纤维细胞

This protocol describes isolation methods, culturing conditions, and characterization of human primary cells with high yield and viability using rapid ...

皮肤含有多种细胞类型，包括角质形成细胞，成纤维细胞和神经施旺细胞。该协议允许分离这些细胞类型中的每一种用于体外实验。从单个供体中分离和建立单个原代细胞培养物可以进行稳健的比较和共培养实验，同时减轻遗传变异的影响。通过该协议，对单个细胞及其相互作用的分析可以分析施旺细胞在皮肤稳态和病理生理学中的作用。这些不同的细胞群可用于研究施旺细胞，成纤维细胞或角质形成细胞，无论是在正常的皮肤生理学，伤口愈合还是皮肤病变状态下组合。Jagadeeshaprasad M G，我实验室的博士后学者将协助我演示该程序。按照标准护理程序获得新生儿包皮后，将切除的包皮置于含有8至10毫升冰冷DMEM基础培养基培养基的微量离心管中，并立即将包皮运送到细胞培养实验室进行细胞分离。用20至25毫升Dulbecco的磷酸盐缓冲盐水或DPBS冲洗包皮两次，含有抗生素和抗真菌剂。然后将洗过的皮肤浸泡在25至30毫升冰冷的DMEM基础培养基中，放入10厘米的组织培养皿中。15分钟后，用手术刀将包皮切开，露出真皮和皮下脂肪组织。使用镊子，手术刀刀片和剪刀去除并丢弃皮下脂肪组织。然后将干净的包皮切成三到五个较小的块，并将皮块在无菌锥形管中的16至20毫升Dispace一个DMEM培养基中孵育。在与Dispace孵育的前30分钟内，通过倒置间歇性地混合锥形管中的皮肤碎片。然后将锥形管置于4摄氏度下16至18小时。第二天取出表皮块，将它们放在干燥的无菌培养皿上，展开每块皮肤，使表皮外层接触到培养皿。从组织边缘开始，使用两组镊子将表皮从真皮上剥离。将分离的表皮碎片加入10厘米培养皿中的5毫升HBS中，在室温下孵育10分钟。然后将HBS溶液收集在50毫升锥形管中，并在锥形管中加入5毫升胰蛋白酶中和缓冲液或T和B。将管子放在一边。在培养皿中的表皮碎片中加入三毫升胰蛋白酶溶液，在37摄氏度下孵育10分钟。孵育后，用镊子轻轻搅拌表皮片段，直到胰蛋白酶角质形成细胞溶液变得浑浊。接下来将胰蛋白酶角质形成细胞溶液加入HBS T和B管中。用胰蛋白酶EDTA溶液溶解未消化的表皮碎片后，向含有胰蛋白酶EDTA溶液的HBS T和B管中加入两毫升FBS，并在四摄氏度下将两者离心约870倍G五分钟。然后吸出上清液并将细胞沉淀重悬于三毫升角质形成细胞完全生长培养基中，然后用无菌的100微米过滤器过滤细胞悬浮液到无菌的50毫升锥形管中。一旦细胞数量和细胞活力被量化，将培养板中的细胞接种到培养板中，以37摄氏度的5%二氧化碳置于培养箱中。两天后，吸出未粘附的细胞，并每隔一天刷新细胞培养基，直到角质形成细胞达到50%至80%汇合度，用于传代或下游实验。用聚-l-赖氨酸或PLL预涂覆T25烧瓶，使用五毫升DPBS，并将预涂好的烧瓶在37摄氏度下放置三小时。随后用DPBS洗涤PLL涂层基体两次。用五毫升DMEM基底培养基洗涤分离的真皮碎片后，用剪刀将真皮切成小块。用五毫升胶原酶在37摄氏度下消化切碎的真皮，持续两个半小时，每30分钟使用移液器尖端轻轻滴定，直到真皮完全解离。消化后，用70微米的细胞过滤器过滤细胞悬浮液，然后用五毫升完整的DMEM培养基稀释过滤的细胞悬浮液，以停止胶原酶的酶活性。接下来，在室温下以870倍G离心细胞悬浮液五分钟，并将细胞轮重悬于两毫升DMEM完全培养基中以分裂细胞。在如上所述重复浓缩过程后，吸出上清液以使用细胞沉淀分离施旺细胞或成纤维细胞。为了分离Schwan细胞，将细胞沉淀重悬于五毫升新鲜DMEM完全培养基中，并量化细胞数量和活力，然后将五毫升重悬的细胞接种在预先编码的T25烧瓶中，密度为4.0倍10到每毫升三个细胞。将烧瓶在37摄氏度和5%二氧化碳下孵育。16小时后，通过显微镜以10倍放大倍率确认细胞粘附。然后除去不粘附的细胞，并用五毫升DPBS洗涤烧瓶三次。接下来，将细胞与5毫升10微摩尔胞嘧啶阿拉伯糖苷在DMEM完全培养基中孵育。24小时后，吸出含有胞嘧啶阿拉伯糖苷的培养基，然后如前所述冲洗烧瓶。用五毫升施万细胞重新填充培养瓶，完成培养基孵育48小时，同时每隔一天刷新培养基，直到施万细胞达到80%汇合度。为了分离成纤维细胞，将收集的沉淀重悬于五毫升成纤维细胞的完整培养基中。在非编码的T25培养瓶中培养细胞24小时，如施万细胞所示。吸出不粘附的细胞并洗涤烧瓶后，将细胞与5毫升新鲜成纤维细胞在37摄氏度和5%二氧化碳下孵育48小时。将分离出的原代细胞置于各细胞培养基中，原代角质形成细胞在第7天达到85%汇合度，并表现出鹅卵石形状的特征性细胞形态。施旺细胞在第五天达到95%汇合，形状为双极或三极。成纤维细胞培养物在第四天达到95%汇合，大多数细胞表现出纺锤形形态。免疫荧光染色证实了细胞培养物中细胞类型特异性蛋白的表达。角质形成细胞对K10和K14蛋白呈阳性。角蛋白免疫荧光和DAPI核染色的合并图像表明细胞培养物的纯度为97.8%。同样，施旺细胞的S100和P75 NTR呈阳性，并且在培养物中的纯度为95.2%。成纤维细胞阳性表达vimentin，但不表达α平滑肌肌肌动蛋白，肌成纤维细胞标志物。培养物的纯度约为97.2%，为了增强Shanse的证据，可以使用预涂的聚-l-赖氨酸皿。为了进一步减轻纤维污染，胞嘧啶阿拉伯糖苷和抗真菌剂在细胞加入短时间后加入。这是一种简单廉价的实验程序，用于从单个人体前皮建立三个原代细胞，如shwann细胞，角质形成细胞和成纤维细胞。该协议非常适合涉及细胞细胞通讯或细胞类型之间串扰的体外实验。

Research

JoVE Journal

Neuroscience

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

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 ...

科研

神经科学

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 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 ...

Isolation and Culture of Endothelial Cells from the Embryonic Forebrain

从胚胎前脑内皮细胞的分离和培养

Embryonic brain endothelial cells can serve as an important tool in the study of angiogenesis and neurovascular development and interactions. The two ...

Isolation, Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

原发性脑多巴胺能神经元的分离，培养和长期维持胚胎脑鼠类

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson&#8217;s diseae. Study of the biological processes ...

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 ...

Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts

人类成人皮肤成纤维细胞诱导神经元高产培养的简单生成

Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue that is easily ...

Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation

密度梯度离心法分离小鼠原发胶质细胞的研究

Microglia, the resident immune cells in the brain, are the first responders to inflammation or injury in the central nervous system. Recent research has ...

Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release

背根神经节分离与原代培养对神经递质释放的研究

Dorsal root ganglia (DRG) contain cell bodies of sensory neurons. This type of neuron is pseudo-unipolar, with two axons that innervate peripheral ...

Primary Culture of Neurons Isolated from Embryonic Mouse Cerebellum

从胚胎小鼠小脑分离的神经元的主要培养

The use of primary cell cultures has become one of the major tools to study the nervous system in vitro. The ultimate goal of using this simplified model ...