SAFETY DATA SHEET.pdf

SPECIFICATION.pdf

使用本产品配制培养基培养的小鼠ES细胞，可用于制备嵌合体小鼠，并确认其种系传递。

·特点 优势

·可用于嵌合体小鼠制备，并确认其种系传递。

·作为血清替代品，用于小鼠ES细胞·人IPS细胞培养。

·不含剧毒物质。

·案例 应用

1、嵌合体小鼠的制备与种系传递Update！！

   将组装HaloTag® cDNA的目标载体导入小鼠ES细胞，用含有Stemsure® Serum Replacement（SSR）的培养基培养。确认载体导入小鼠ES细胞后，将其注入到CR小鼠的胚盘胞，并移植至ICR小鼠的子言。得到的嵌合体小鼠与C57BL/6J小鼠交配，获得F1小鼠。

（1）嵌合体小鼠

   收集嵌合体小鼠的血液，用TRITC（四甲基若丹明异硫氰酸盐）对HaloTag® 蛋白质染色。发现小鼠ES细胞源的HaloTag 的血液被染色，可确认为嵌合化。

（2）F1小鼠

   F1小鼠的血液发现HaloTag® 存在，可确认用SSR培养基培养的ES细胞嵌合小鼠能够正常存活。

2、人iPS细胞201B7的培养

（1）克隆形态与ALP染色

    已确认可用StemSure®  Serum Replacement（SSR）培养人iPS细胞201B7株。ALP染色呈阳性。

   <培养基组成>

D-MEM/Ham's F-12+20%SSR +2 mmol/l L-Glutamine +1×MEM Non-essential Amino Acids +0.1

mmol/l StemSure® 2-Mercaptoethanol +1xPenicillin-Streptomycin +5 ng/ml bFGF

（2）未分化标记的发现

   使用StemSure® Serum Replacement（SSR）培养的人iPS细胞201B7株，确认各种未分化标记（Sox2，Oct3/4，SSEA-3，SSEA-4，Tra-1-81）呈阳性。

3、小鼠ES细胞D3的培养

（1）细胞形态与ALP染色

   使用StemSure® Serum Replacement（SSR）培养小鼠ES细胞D3，能观察到小鼠ES细胞群特有的光泽，ALP染色呈阳性。

（2）细胞增殖曲线

   用不同培养基培养小鼠ES细胞D3进行14继代培养，比较细胞增殖。

   如下图所示StemSure®系列与A公司有同等的性能。

<StemSure®系列培养基组成>

StemSure®D-MEM+15%SSR +2 mmol/l L-Glutamine +1xMEM Non-essentialAmino Acids +0.1

mmol/lStemSure® 2-Mercaptoethanol+1×Penicillin-Streptomycin +1，000 units/ml StemSure®LIF

（使用胶原包被的12孔板）

（3）细胞群倍加数*

   使用不同培养基对小鼠ES细胞D3进行继代培养，检查培养天数与细胞群倍加数的关系。

   下图显示StemSure®系列与A公司产品的细胞群倍加数相同。

*细胞群倍加数cell population doubling level，缩写PDL，指从培养开始至现在细胞群的倍增次数。

<培养基组成>

StemSure® D-MEM+15%SSR+2 mmol/l L-Glutamine +1xMEM Non-essentialAmino Acids +0.1

mmol/l StemSure® 2-Mercaptoethanol +1×Penicillin-Streptomycin +1，000 units/ml StemSure® LIF

（使用胶原包被的12孔板）

（4）未分化标记的检测

    使用StemSure®系列对小鼠ES细胞D3进行5代继代培养，各种未分化标记（Nanog，Oct3/4，Sox2，SSEA-1）呈阳性。

<培养基组成>

StemSure® D-MEM+15%SSR+2 mmol/l L-Glutamine +1×MEM Non-essential Amino Acids +0.1

mmol/l StemSure® 2-Mercaptoethanol+1xPenicillin-Streptomycin +1，000 units/ml StemSure® LIF

4、畸形瘤的形成

   用StemSure®系列对小鼠ES细胞D3进行继代培养。对免疫不全小鼠的皮下注射培养后细胞。在形成皮下畸形瘤，其内部检测出神经组织（外胚叶源）、软骨组织（中胚叶源）、管腔结构纤毛上皮（内胚叶源）。

<培养基组成>

StemSure® D-MEM+15%SSR+2 mmol/l L-Glutamine +1×MEM Non-essential Amino Acids +0.1

mmol/l StemSure® 2-Mercaptoethanol+1xPenicillin-Streptomycin +1，000 units/ml StemSure® LIF

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