Name: passaging hues human embryonic stem cell-lines with trypsin.
Uploaded: 2006-10-12T00:00:00
Description: Watch this Scientific Journal Video about Passaging HuES Human Embryonic Stem Cell-lines with Trypsin. at JoVE.com

一般我们建议解冻在一个给定的的时间不超过一个样本，以确保容易处理。处理程序应注意不要长时间留在室温和低二氧化碳的文化。所有活细胞的离心是在室温下5分钟在500-600 XG。 MEFs（小鼠饲养层细胞） <ol><li>预温MEF的媒体至37 ° C。删除MEF的小瓶从-80 ° C，并立即淹没在37℃水浴试管底部的一半。细胞解冻80％（左小冻结部分）之前，需要大约30-45秒。...

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在这个视频中，我们证明我们定期通过色调如何在我们的实验室中的人类胚胎干细胞系。高效和定期的分裂和传代是保持一个健康的人类胚胎干细胞系的关键。胰蛋白酶介导的通过是一个色调细胞系的显着优势，但是，重要的是不要过分trypsinize文化或有一个单细胞的过程中再platting大的比重。

色调媒体651.5毫升KO - DMEM培养液500毫升PenStrep 6.5毫升GlutaMAXTM 6.5毫升NEAA 6.5毫升2 - 巯基乙醇0.5ul KO血清替代65毫升Plasmanate bFGF2（10毫微克/毫升决赛）0.65毫升65毫升

passaging hues human embryonic stem cell-lines with trypsin.

在这个视频中，我们展示了我们的实验室常规通道与胰蛋白酶色调人类胚胎干细胞系。人类胚胎干细胞是细胞培养的产物，并倾向于收购与高人口倍增的异常核型。正确的传代是保持一个健康的，未分化，核型正常的色调的人类胚胎干细胞培养的关键。首先，扩大文化是在PBS洗涤，以去除残留的媒体和的细胞碎片，然后将细胞覆盖一个温暖的0.05％胰蛋白酶EDTA的最小量。胰蛋白酶是留给长达五分钟的细胞，然后细胞被轻轻抛下一个2ML血清吸管。细胞悬液与色调媒体的大量收集和混合，然后收集细胞，轻轻离心。胰蛋白酶灭活媒体混合物被删除，细胞悬浮预热色调媒体。一个适当的分流比计算（一般为1:10至1:20），和细胞到1-2天老照射的小鼠胚胎成纤维细胞饲养层细胞单层板重新镀金。新种子的色调培养板，左为48小时不受干扰，然后媒体是每天更换其后。重要的是不要trpsinize单细胞悬液，因为这增加了引进核型异常的风险。

Watch this Scientific Journal Video about Passaging HuES Human Embryonic Stem Cell-lines with Trypsin. at JoVE.com

Passaging HuES Human Embryonic Stem Cell-lines with Trypsin.

在这个视频中，我们展示了我们的实验室常规通道与胰蛋白酶色调人类胚胎干细胞系。

人类胚胎干细胞传代的色调线与胰蛋白酶

In this video we demonstrate how our lab routinely passages HuES human embryonic stem cell lines with trypsin.  Human embryonic stem cells are artifacts ...

Research

JoVE Journal

Biology

Propagation of Human Embryonic Stem (ES) Cells

Propagation of Human Embryonic Stem ES Cells

人类胚胎干细胞（ES细胞）的繁殖

科研

生物学

Passaging Human Neural Stem Cells

传代人类神经干细胞

The ability to manipulate human neural stem/precursor cells (hNSPCs) in vitro provides a means to investigate their utility as cell transplants for ...

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 ...

Probing for Mitochondrial Complex Activity in Human Embryonic Stem Cells

在人类胚胎干细胞的线粒体复杂的活动初探

Mitochondria are cytoplasmic organelles that have a primary role in cellular metabolism and homeostasis, regulation of the cell signaling network, and ...

Chromatin Immunoprecipitation from Human Embryonic Stem Cells

从人类胚胎干细胞的染色质免疫沉淀

The functional and structural complexity of the myriad of cells in metazoan organisms arises from a small number of stem cells. Stem cells are ...

Culture and Maintenance of Human Embryonic Stem Cells  

文化和维护人类胚胎干细胞

Human embryonic stem (hES) cells must be monitored and cared for in order to maintain healthy, undifferentiated cultures. At minimum, the cultures must be ...

Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping

为核型分析人类胚胎干细胞的染色体蔓延制备

Although human embryonic stem cells (hESC) have been shown to present a stable diploid karyotype 1, many studies have reported that depending on culture ...

Freezing and Thawing Human Embryonic Stem Cells

人类胚胎干细胞冷冻和解冻

Since James Thomson et al developed a technique in 1998 to isolate and grow hES in culture, freezing cells for later use and thawing and expanding cells ...

Video Bioinformatics Analysis of Human Embryonic Stem Cell Colony Growth

视频人类胚胎干细胞克隆生长的生物信息学分析

Because video data are complex and are comprised of many images, mining information from video material is difficult to do without the aid of computer ...

Differentiation of Embryonic Stem Cells into Oligodendrocyte Precursors

胚胎干细胞分化成少突胶质前体

Oligodendrocytes are the myelinating cells of the central nervous system. For regenerative cell therapy in demyelinating diseases, there is significant ...