Name: isolation and derivation of mouse embryonic germinal cells
Uploaded: 2009-10-22T14:00:00
Description: Watch this Scientific Journal Video about Isolation and Derivation of Mouse Embryonic Germinal Cells at JoVE.com

作者想承认了宝贵的帮助：尼尔第一手稿的编辑和Kourtney威尔金森博士;露西信通博士，博士Brigit Willeford和迈克Basett在密歇根州立大学ALAC的认可鼠标设施，培训和动物服务的援助;德韦恩明智博士为他的援助显微镜和图像采集;塞萨尔蒙罗伊，汉娜Swoope，协助他们在密歇根州立大学与视频生产和Bobbie赫德尔斯顿。这项研究是由研究机构和密西西比州立大学生物科学系办公室。

第1部分：孕鼠剖腹<ol><li>使用颈椎脱位，安乐死在10.5-11.5 DPC怀孕C57BL6J雌性小鼠。 </li><li>抗菌肥皂与清洁腹部，然后，刮胡子。 </li><li>剃须后，用生理盐水溶液冲洗腹部。 </li><li>然后，用无菌纱布擦干腹部。 </li><li>盖鼠标与一个新的无菌领域。 </li><li>制作腹切口，使用镊子和解剖剪刀。 </li><li>识别和清除腹腔整个子宫。小鼠胚胎将子宫内可见。 </li><li>转移到培养皿中弥漫着的D - PBS的子宫，?...

<ol>
	<li>DeFelici, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Cell Biology: A laboratory Handbook. 1, 73-85 (1998).</li><li>Donovan, P. J., De Miguel, 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=Turning+Germ+Cells+into+Stem+Cells.">Turning Germ Cells into Stem Cells.</a> Current Opinion in Genetics and Development. 13, 463-471 (2003).</li><li>Labosky, P. A., Hogan, B. L. M., Sharp, P. T., Mason, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Part+12:+Mouse+Primordial+Germ+Cells:+isolation+and+in+vitro+culture.">Part 12: Mouse Primordial Germ Cells: isolation and in vitro culture.</a> Molecular embryology: methods and protocols. 97, (1999).</li><li>Mc Laren, A., Southee, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Entry+of+Mouse+Embryonic+Germ+Cells+into+Meiosis.">Entry of Mouse Embryonic Germ Cells into Meiosis.</a> Developmental Biology. 187, 107-113 (1997).</li><li>Yoshimizu, T., Obinata, M., Matsui, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stage-specific+tissue+and+cell+interactions+play+key+roles+in+mouse+germ+cell+specification.">Stage-specific tissue and cell interactions play key roles in mouse germ cell specification.</a> Development. 128, 481-490 (2001).</li><li>Zhang, J., Hhvorostov, I., Teitell, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+MEFs+to+Matrigel+I:+Passaging+hESCs+in+the+Presence+of+MEFs.">From MEFs to Matrigel I: Passaging hESCs in the Presence of MEFs.</a> J Vis Exp. , (2008).</li></ol>

钳第14号和15日，手术刀35毫米，解剖剪刀，手术领域，抗菌肥皂，盐水0.9％，纱布，冰泡沫容器，刀片，手术刀35毫米，4号和5级罚款钳，细戏弄针处理，过滤纸，然后在拉玻璃吸液管。<br /<br /&gt;设备：解剖立体显微镜，光源肖特Fostec，倒置显微镜，微量移液器10和200ul，离心机，生物安全柜。

isolation and derivation of mouse embryonic germinal cells

胚胎生殖细胞的能力（例如），以区分在早期发育阶段，是一个完美的模型，以解决我们的假设，对癌症和不育症的原始生殖细胞（PGCS），后来进入配子。该协议显示了如何在10.5-11.5天coitum（DPC）的小鼠胚胎来自发展中国家性腺的原始生殖细胞分离。发展中国家从小鼠胚胎性腺脊（C57BL6J）分离机械破碎与胶原酶，然后镀上了一个小鼠胚胎成纤维细胞饲养层（MEF - CF1）以前分裂与丝裂霉素C灭活淘汰赛媒体的存在，并辅以白血病抑制，因子（LIF），碱性成纤维细胞生长因子（bFGF），干细胞因子（SCF）。使用这些优化方法盈科鉴定，隔离，并建立培养条件允许超过40天，EG细胞的长期培养。胚胎生发细胞株显示胚胎的表型多能状态的常用标记和表达。文化的生发细胞的分离和衍生提供了一个工具，以了解体外他们的发展提供了机会，在遗传和表观遗传一级监察累积损伤后接触氧化应激。

Watch this Scientific Journal Video about Isolation and Derivation of Mouse Embryonic Germinal Cells at JoVE.com

Isolation and Derivation of Mouse Embryonic Germinal Cells

胚胎生殖细胞分化成原始生殖细胞在早期发展阶段的能力，是一个完美的模型，以解决我们的假设，对癌症和不育症。该协议显示了如何从发展中国家性腺的原始生殖细胞中分离出来后10.5-11.5天coitum小鼠胚胎。

小鼠胚胎生殖细胞的分离和推导

The ability of embryonic germinal cells (EG) to differentiate into primordial germinal cells (PGCs) and later into gametes during early developmental ...

Research

JoVE Journal

Biology

Derivation of Hematopoietic Stem Cells from Murine Embryonic Stem Cells

从小鼠胚胎干细胞的造血干细胞的推导

A stem cell is defined as a cell with the capacity to both self-renew and generate multiple differentiated progeny.  Embryonic stem cells (ESC) are ...

科研

生物学

Derivation of Human Embryonic Stem Cells by Immunosurgery

人类胚胎干细胞的推导Immunosurgery

The ability of human embryonic stem cells to self-renew and differentiate into all cell types of 
the body suggests that they hold great promise for both ...

Derivation of Mouse Trophoblast Stem Cells from Blastocysts

鼠标滋养层干细胞从胚泡的推导

Specification of the trophectoderm is one of the earliest differentiation events of mammalian development. The trophoblast lineage derived from the ...

Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells

成人和胚胎骨骼的肌肉Microexplant文化和骨骼肌肉干细胞的分离

Cultured embryonic and adult skeletal muscle cells have a number of different uses. The micro-dissected explants technique described in this chapter is a ...

Isolation and Culture of Avian Embryonic Valvular Progenitor Cells

禽流感胚胎瓣膜祖细胞的分离和培养

Proper formation and function of embryonic heart valves is critical for developmental progression. The early embryonic heart is a U-shaped tube of ...

Isolation and Culture of Cells from the Nephrogenic Zone of the Embryonic Mouse Kidney

从小鼠胚胎肾肾区细胞的分离和培养

Embryonic development of the kidney has been extensively studied both as a model for epithelial-mesenchymal interaction in organogenesis and  to gain ...

Efficient Derivation of Human Cardiac Precursors and Cardiomyocytes from Pluripotent Human Embryonic Stem Cells with Small Molecule Induction

高效的推导人类心脏的前体和从多能干的小分子诱导人类胚胎干细胞的心肌细胞

To date, the lack of a suitable human cardiac cell source has been the major setback in regenerating the human myocardium, either by cell-based ...

Preparation of Mouse Embryonic Fibroblast Cells Suitable for Culturing Human Embryonic and Induced Pluripotent Stem Cells

适用于培养人类胚胎和诱导多能干细胞的小鼠胚胎成纤维细胞的制备

In general, human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs)1 can be cultured under variable conditions. However, it ...

Isolation of Embryonic Ventricular Endothelial Cells

胚胎心室血管内皮细胞的分离

Cell culture has greatly enhanced our ability to assess individual populations of cells under myriad culture conditions. While immortalized cell lines ...

Isolation and Culture of Mouse Primary Pancreatic Acinar Cells

小鼠原代胰腺腺泡细胞的分离和培养

This protocol permits rapid isolation (in less than 1 hr) of murine pancreatic acini, making it possible to maintain them in culture for more than one ...