CTS™ CytoTune™-iPS 2.1 Sendai Reprogramming Kit - FAQs

View additional product information for CTS™ CytoTune™-iPS 2.1 Sendai Reprogramming Kit - FAQs (A34546)

15 product FAQs found

TaqMan仙台病毒基因检测Assay是否兼容所有版本的CytoTune试剂盒？

TaqMan仙台病毒基因检测Assay和CytoTune 2.0和CytoTune 2.1版本都兼容，但有一个例外。CytoTune 2.0试剂盒包含一个表达c-Myc的载体，而CytoTune 2.1试剂盒包含的是表达L-Myc的载体。因此，对每种试剂盒应使用与之相对应的Assay(例如，SEVCMYC用于CytoTune 2.0, SEVLMYC用于CytoTune 2.1)。所有其它Assay对这两种试剂盒都是兼容的。

我能区分基因（SOX2，Kfl4，cMyc和Oct2）的内源序列和外源序列吗？你们是否提供能区分这些基因内源序列与外源序列的引物？

我们没有用于终点法RT-PCR的这种引物。然而，我们确实为每一个外源性重编程基因提供了经过验证的TaqMan基因表达Assay (https://www.thermofisher.com/order/genome-database/browse/gene-expression/keyword/sendai+taqman)。这些Assay被设计用来专门检测仙台病毒载体上发现的外源基因的表达，不会与这些基因的内源性表达发生交叉反应。此外，这些Assay也可被用于检测每种CytoTune仙台病毒载体的存在，以确定这些载体是否从已建立的ipsc中清除了。  

我该如何确认我的细胞已经被仙台病毒成功转导？

您可通过多种方法确认细胞中仙台病毒的存在，包括使用抗仙台病毒的抗体进行染色，或使用CytoTune-iPS 2.0仙台病毒重编程试剂盒用户手册 (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cytotune_ips_2_0_sendai_reprog_kit_man.pdf)第50页上所列的引物对进行终点法RT-PCR检测，或使用TaqMan Sendai Gene Expression assays（也列在说明书第50页上）进行实时RT-PCR检测。

What is CTS?

The Gibco Cell Therapy Systems (CTS) portfolio of cell and gene therapy products are GMP manufactured, safety tested, and backed by regulatory documentation to support your transition from discovery through clinical and commercial manufacturing. Through our CTS solutions, we are committed to helping customers streamline therapeutic development, minimize risk, and ease the burden on their quality systems. Learn more here.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How many transductions can I perform with one CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit?

The CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit is sufficient to reprogram a minimum of six wells in a six-well plate. One well has a surface area of approximately 10 cm2.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Are the reprogramming steps in the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit different compared to CytoTune-iPS 2.0 Sendai Reprogramming Kit?

No, the reprogramming steps are in principle the same but it is important to consider the difference in titer between the two kits when transducing cells at certain MOIs.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Why is the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit so much more expensive than the CytoTune-iPS 2.0 Sendai Reprogramming Kit?

It is more expensive because of its label use statement: For research use and non-commercial manufacturing of cell based products for clinical research. CAUTION: Not intended for direct administration into animals or humans. The kit is manufactured according to GMP requirements and the drug master file is available for submission to FDA upon customer request, for use in clinical research.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Why does the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit contain L-Myc virus instead of c-Myc virus?

The L-Myc virus was chosen over the c-Myc virus because it is considered superior from a safety perspective, for clinical applications (L-myc is reported to have lower transformation/oncogenic potential than c-myc).

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

What is the difference between the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit and the CytoTune-iPS 2.0 Sendai Reprogramming Kit?

The CTS CytoTune-iPS 2.1 Sendai vector formulation does not contain the bovine serum albumin carrier protein. Additionally, the preparation contains no animal-derived components at the primary component level. Each virus (i.e., KOS, L-Myc, and Kfl4) comes in a volume of 200 µL instead of 100 µL. The titer of the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit can range from 0.5 x 10E8 to 2.0 x 10E8 virus particles per mL whereas the titer of the CytoTune-iPS 2.0 Sendai Reprogramming Kit is typically around 1.0 x 10E8 virus particles per mL.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

What do I need to know about the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit if I am interested in reprogramming cells, and my project could be part of clinical research in the near future?

The kit is manufactured according to GMP requirements and the drug master file is available for submission to FDA upon customer request.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

What is the label use statement for the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit?

The label use statement is listed on the Certificate of Analysis (COA) and is as foll:

For research use and non-commercial manufacturing of cell based products for clinical research. CAUTION: Not intended for direct administration into animals or humans.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Now that you offer a CytoTune 2.1 kit, has the CytoTune 2.0 kit been discontinued?

No, we still offer the CytoTune 2.0 kit.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Are the TaqMan Sendai Gene Expression assays compatible with all versions of CytoTune?

The TaqMan Sendai Gene Expression Assays are compatible with both CytoTune 2.0 and CytoTune 2.1, with one exception. The CytoTune 2.0 kit contains a vector which expresses c-Myc, while the CytoTune 2.1 kit contains a vector which expresses L-Myc. As such, the relevant assays should be used in conjunction with each kit (e.g. SEVCYMC for CytoTune 2.0, and SEVLMYC for CytoTune 2.1). All other assays are compatible with both kits.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I distinguish between the endogenous and exogenous expression of the genes (SOX2, Kfl4, cMyc, and Oct2)? Do you offer primers that are able to distinguish between endogenous and exogenous expression of these genes?

We do not have such primers for use in endpoint RT-PCR. However, we do offer validated TaqMan Gene Expression assays for each of the exogenous reprogramming genes (https://www.thermofisher.com/order/genome-database/browse/gene-expression/keyword/sendai+taqman). These assays are designed to specifically detect only expression of the exogenous genes found on the Sendai vectors, and will not cross-react with expression of the endogenous versions of these genes. Additionally, these assays can be used to detect the presence of each of the CytoTune Sendai vectors, for determination of clearance of the vectors from established iPSCs.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How do I confirm that my cells were transduced with the Sendai virus?

You can confirm the presence of the Sendai virus in your cells by several methods, including staining with anti-Sendai virus antibody, performing endpoint RT-PCR using the primer sets listed on page 50 in the CytoTune-iPS 2.0 Sendai Reprogramming Kit User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cytotune_ips_2_0_sendai_reprog_kit_man.pdf), or real-time RT-PCR using the TaqMan Sendai Gene Expression assays, also listed on page 50 in the CytoTune-iPS 2.0 Sendai Reprogramming Kit User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cytotune_ips_2_0_sendai_reprog_kit_man.pdf).

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.