MicroRNA-Enhanced mRNA Reprogramming System
产品信息
Product Name |
Cat. No. |
Size |
Storage |
Stemgent® mRNA Reprogramming Kit |
00-0071 |
|
|
Kit Contents: |
|
|
mRNA Reprogramming Factors Set: hOKSM |
7×200 μl |
-70℃ |
B18R Recombinant Protein (0.5 mg/ml) |
80 μl |
-70℃ |
Pluriton™ Reprogramming Supplement (2500×) |
0.2 ml |
-70℃ |
Pluriton™ Reprogramming Medium |
500 ml |
-20℃ |
Stemgent® microRNA Booster Kit |
00-0073 |
|
|
Kit Contents: |
|
|
microRNA Reprogramming Cocktail |
2×35 μl |
-70℃ |
B18R Recombinant Protein (0.5 mg/ml) |
80 μl |
-70℃ |
Stemgent® Stemfect™ RNA Transfection Kit |
00-0069 |
|
|
Kit Contents: |
|
|
Stemfect™ RNA Transfection Reagent |
750 μl |
4℃ |
Stemfect™ Buffer |
35 ml |
4℃ |
※ 上述三个试剂盒联合使用,能够满足9-10个Well(6-well plate format)内靶细胞的重编程。
产品特点
1.快速 Fast
Stemgent® MicroRNA-Enhanced mRNA Reprogramming System只需2周即可将靶细胞重编程为iPS细胞,与仙台病毒(Sendai Virus)、非整合型DNA载体(Episomal DNA Vectors)等传统iPS重编程方法相比,缩短实验周期超过50%以上(Figure. 1)。

Figure 1. Stemgent® mRNA重编程系统、仙台病毒依赖的重编程系统以及游离型DNA载体依赖的重编程系统重编程时程比较。
简便 Easy
与病毒、DNA载体依赖的iPS重编程手段不同,使用Stemgent® MicroRNA-Enhanced mRNA Reprogramming System对靶细胞进行重编程,生成的iPS细胞无需进行复杂耗时的下游筛选程序,只需简简单单6个步骤,即可轻松构建iPS细胞系。
|
Description |
Day |
Step 1 |
Material Preparation |
Prior to starting |
Step 2 |
Plate Cells |
0 |
Step 3 |
Transfect Cells |
1 to 12 |
Step 4 |
Identify Cells |
13-14 |
Step 5 |
Pick and Passage iPS Cell Colonies |
15-16 |
Step 6 |
Maintain iPS Cell Cultures |
16+ |

注:因所用细胞类型及实验条件不同,上述时间轴可能略有不同,仅供参考。

Figure 2. Stemgent® MicroRNA-Enhanced mRNA Reprogramming System重编程过程时间表、显微镜下观察到的重编程过程中靶细胞形态变化。第12天,形成的iPS克隆表达多能型Marker TRA-1-81。
高效 Efficient
病毒、DNA载体等依赖的iPS重编程技术,其重编程效率介于0.00001%到1%之间不等,而Stemgent® MicroRNA-Enhanced mRNA Reprogramming System的重编程效率远远大于1%,极大地提高了iPS克隆的产出率。
表1. 不同类型iPS重编程手段效率比较
Reprogramming Method |
Efficiency |
Integrating |
Screening |
RNA |
>1% |
No |
No |
仙台病毒(Sendai virus) |
0.01-1% |
No |
Yes |
DNA载体(Episomal/Minicircle) |
0.00% |
Possible |
Yes |
慢病毒(Lentivirus) |
0.001-0.01% |
Yes |
Yes |
腺病毒(Adenovirus) |
0.0001-0.001% |
Possible |
Yes |
Protein |
0.00% |
No |
No |
安全 Safe
某些iPS研究领域,例如iPS临床研究、治疗应用等,对iPS细胞的安全性提出了极高的要求。使用Stemgent® MicroRNA-Enhanced mRNA Reprogramming System重编程靶细胞,无需担心构建的iPS细胞系可能含有病毒载体残留,亦无需担心无关的基因整合进宿主基因组,是目前适用于临床研究及治疗的最理想选择。
而以仙台病毒(Sendai Virus)为载体的iPS重编程手段,其病毒基因组虽然不会整合进宿主染色体,仙台病毒本身也不具备复制能力,但是为了构建真正意义上的virus-free的iPS细胞系,通常需要经过一个10-20代、耗时费力的筛选过程,以确保最终得到的iPS细胞系无病毒载体残留。以慢病毒(Lentivirus)体的iPS重编程手段,病毒基因组能够永久整合进宿主基因组,会给后续的iPS细胞分析带来干扰。

高性价比 Cost Effective
以仙台病毒为载体的iPS重编程试剂盒非常昂贵,综合重编程效率、时间及劳动力成本、安全性等多重考虑,Stemgent® MicroRNA-Enhanced mRNA Reprogramming System无疑是性价比的上乘之选。
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产品订购,请电询:4008-217-320