- 产品特性
- 相关资料
- Q&A
- 参考文献
细胞培养基质 层粘连蛋白511
iMatrix-511
◆什么是层粘连蛋白511?
层粘连蛋白是存在于动物基底膜的一种细胞外基质,已知其与细胞粘附和增殖息息相关。本产品是与层粘连蛋白 511-E8 片段有同一序列的重组蛋白,是可以促进各种细胞粘附和伸展的培养基质。
大阪大学和京都大学共同研究开发,本产品已证明在操作难度非常大的人 iPS 细胞和人 ES 细胞培养中,也可以安全且高效地进行细胞培养。
● 细胞培养的准备非常简单!
● 可用于多种类型的细胞培养!
● 无论分离细胞的状态如何,可实现细胞的高生存率和细胞增殖的高效性!
● 重组蛋白(CHO-S细胞来源),所以混入杂质的危险性低!
● 溶液型试剂,无需溶解,稀释后可直接使用
层粘连蛋白511 是由 α5 链、β1 链和 γ1 链组成的层粘连蛋白。层粘连蛋白 511-E8 是层粘连蛋白片段,但其具有与层粘连蛋白全长分子相同的 α6β1 整合蛋白连接功能。
本产品是 Nippi 根据大阪大学和京都大学的专利技术生产贩卖的。
◆使用方法
用 PBS(-)稀释本产品,按照 0.1~0.5 μg/cm2 加入细胞培养器皿中。
※由于细胞种类和细胞株的不同、使用的培养基种类不同,添加的最适剂量会有差异,初次使用时,按照 0.5 μg/cm2 添加培养容器中,逐渐
※调整至最佳使用浓度。
↓
室温下孵育 3 小时,然后去掉溶液。
↓
添加细胞和培养液,进行细胞培养。
● 培养 ES/iPS 细胞时,可进行无饲养层和单细胞继代培养
◆使用案例
使用本产品对表皮细胞培养 0.5 小时,对血管内皮细胞培养 1 小时。
(a) 表皮细胞,培养 0.5 小时
左(无涂层):大部分细胞未贴壁。
右(iMatrix):多数细胞贴壁并成伸展状态。
(b) 血管内皮细胞,培养 1 小时
左(无涂层):有贴壁的细胞,但大部分多为圆形。
右(iMatrix):较少观察到圆形细胞,几乎所有的细胞表现出了很好的伸展性。
实验人员: (株) Nippi BioMatrix 研究所 藤崎
◆iMatrix-511 与 iMatrix-511 silk 的区别
iMatrix-511 |
iMatrix-511 silk |
|
生产系统 |
转基因 CHO-S 细胞 |
转基因蚕生产系统 |
提纯材料 |
CHO-S 细胞培养上清 |
蚕蛹蛋白 |
产品等级 |
实验研究用* *有临床用级别 |
实验研究用 |
导入基因 |
人层粘连蛋白 511-E8 片段 |
|
纯度 |
95% 以上 |
|
浓度 |
0.5 mg/mL |
|
解离常数 |
10 nM 以下 |
|
使用期限 |
生产后 2 年内 |
|
iPS 细胞培养能力 |
添加 0.5 μg/cm2 到培养容器中,可用于 iPS 细胞的维持培养 |
◆产品列表
产品编号 |
生产商编号 |
产品名称 |
包装 |
385-07361 |
892011 |
iMatrix-511 solution (0.5 mg/mL) 层粘连蛋白511-E8片段, 溶液(0.5 mg/mL) |
175 µg×2(350 µL×2) |
381-07363 |
892012 |
175 µg×6(350 µL×6) |
※ 本页面产品仅供研究用,研究以外不可使用。
点击此处,选择页面中的”iMatrix™ Calculator“计算实验中 iMatrix-511 使用量
点击此处进一步获取文献应用实例
点击此处下载产品宣传页
iMatrix-511,iMatrix-511 silk
Q1. 产品是以什么样的状态销售的?
A1. 产品是以液态销售的。一支试管里密封装有 0.5 mg/mL 浓度的 175 μg 的细胞培养外基质(层粘连蛋白 511-E8 片段)。
※iMatrix-511 的冻干产品已于2015年3月停止生产。
Q2. 产品的存储条件和有效期是什么?
A2. 产品的储存条件为,冷藏保存在 2-15°C。(推荐 4°C)
A2. 产品的有效期,请参考下表。
产品 |
保质期 |
iMatrix-511 |
自生产后两年内 |
iMatrix-511silk |
自生产后两年内 |
iMatrix-511MG |
自生产后两年内 |
※具体的有效期详见产品外包装。
Q3. 可以冷冻保存吗?
A3. 不可以冷冻保存。
Q4. 产品的纯度是多少?
A4. 纯度为 95% 以上。
Q5. 培养 hES/hiPS 细胞时使用什么培养基最好?
A5. 宫崎等的论文(Nature communications, 3(1236), 1-10, 2012)、(Scientific Reports, 7, 41165, 2017)中,使用了以下的培养基。
・mTeSR1,TeSR2,TeSR-E8 (STEMCELL Technologies)
・Stem Pro hESC SFM (Thermo Fisher Scientific)
・StemFit AK03 (Ajinomoto)
中川等的论文(Scientific Reports, 4(3594), 1-7, 2014)中使用了以下的培养基。
・StemFit (Ajinomoto)
文献中使用的培养基都出现了良好的结果。
Q6. 培养 iPS 细胞时,最佳的涂层浓度是多少?
A6. 最佳的涂层浓度根据细胞株的不同也会有所不同。
A6. 最初请从浓度 0.5 μg/cm2 开始尝试,请根据您使用的细胞株在浓度 0.1~1.5 μg/ cm2 之间考虑。
A6. 另外,还有文献报道了新的不以涂层包被的添加法。
A6. Miyazaki et al. Scientific Reports, 7, 41165, (2017)
Q7. 请教我使用 iMatrix-511 时,培养 iPS 细胞的步骤。
A7. 使用 iMatrix-511 时,ES/iPS 细胞的扩增培养步骤,传代操作,请参考以下的链接。
(扩增培养步骤 )
(传代操作的视频)
Q8. 培养 iPS 细胞时,需要 Rock Inhibitor(Y-27632)吗?
A8. 在中川等的论文(Scientific Reports, 4(3594), 2014)中,介绍了只有在传代时添加 Rock Inhibitor,更换培养基的时候不需使用。
Q9. 培养 iPS 细胞时可以单细胞传代吗?
A9. 可以。
※使用 iMatrix-511 时,ES/iPS 细胞的扩增培养步骤,传代操作,请参考以下的链接。
(扩增培养步骤 )
(传代操作的视频)
也可以参考中川等的论文(Scientific Reports, 4(3594), 2014)。
Q10. 传代时使用什么细胞分离液?
A10. 可使用胰蛋白酶。
※使用 iMatrix-511 时,ES/iPS 细胞的扩增培养步骤,传代操作,请参考以下的链接。
(扩增培养步骤)
(传代操作的视频)
也可以参考中川等的论文(Scientific Reports, 4(3594), 2014)。
Q11. 可以使用小鼠的 iPS 细胞吗?
A11. 由于没有小鼠 iPS 细胞的培养数据,所以无法回答。
Q12. 这个产品与基质胶有什么不同?
A12. 基质胶含有小鼠EHS肉瘤来源的层粘连蛋白-111。另外还含有层粘连蛋白以外的分子。
A12. iMatrix-511 是将在 CHO-S 细胞中表达的层粘连蛋白 511-E8 片段高度纯化后的重组蛋白。
A12.iMatrix-511silk 是从蚕结的茧中高纯度纯化层粘连蛋白 511-E8 片段的重组蛋白。
A12.已知人ES细胞和 iPS 细胞是通过细胞膜受体(特别是 α6β1整联蛋白)粘附于层粘连蛋白-511。
A12.已知人ES细胞和 iPS 细胞对层粘连蛋白-511 具有高粘附活性。这使得用 iMatrix-511/iMatrix-511silk 可以使 iPS 细胞在单细胞状态下传代。
A12.在宫崎等的论文中(Nature communications, 3(1236), 1-10, 201),5 次传代后(30 天后)的细胞数扩增效率约为基质胶的 200 倍。
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|
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||
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|
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||
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|
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