The benefits of working with iPSC Reprogramming Services

A major advantage of working with human iPSCs, compared to pluripotent embryonic stem cells, is that these cells have a known phenotype, unlike pluripotent cells from embryos in which the phenotype is unknown.

Similar to human embryonic stem cells (hESCs), iPSCs can be generated and used as a cellular tool to unravel mechanisms of human development and to model diseases in a manner not possible before given that many of these diseases do not occur until later in life. iPSCs also provide a human model system, which is important to consider as certain disease models such as Alzheimer’s disease cannot be accurately replicated in animal models.

Besides these fundamental aspects of human biology and physiology that are revealed using iPSC or iPSC-derived cells, these cells hold an immense potential for cell-based therapies, and for the discovery of new or personalized pharmacological treatments for many disorders. An additional advantage is that many of the ethical concerns around the derivation of hESCs are not applicable to the derivation of iPSCs. 

What are the advantages of outsourcing the reprogramming of adult cells into iPSCs?

1. Our iPSC Reprogramming experts are your experts

One of the key advantages of using an iPSC reprogramming services, such as those offered by Sampled, is the convenience it offers. Before even working with the cells, researching the protocols, best practices and reagents required for efficient and reliable iPSC reprogramming can be a time-consuming and costly process.

By outsourcing this work to a service provider such as Sampled, researchers can access the latest reprogramming iPSC protocols and technologies, which ensures a higher level of efficiency and accuracy in the reprogramming process. We know that reprogramming protocols are long and laborious, and that one simple error can set back researchers by weeks. By outsourcing cell reprogramming, you can stay one step ahead by focusing on the work that matters most and not wasting any time on experiments that fail due to insufficient expertise or knowledge.

2. Quality control that is tailored to meet your needs

By outsourcing iPSC reprogramming services, you are able to tailor the quality control tests in order to get exactly what you need. At Sampled our human iPSC generation service come with several quality control tests as standard. Every cell undergoes rigorous testing to ensure their quality and consistency:

• Sterility testing including mycoplasma
• Cell identity confirmation using 96 SNP Fluidigm Panel
• Post thaw viability
• Expression of stemness markers through FACS analysis of the dual expression of Oct4 and Tra-1-60

This gives researchers confidence in the cells they are using and provides them with a reliable source of cells for their research. Furthermore, we offer bespoke quality control options that include:

• Analysis of pluripotency through Scorecard: Demonstration of pluripotency using in vitro assays such as embryoid body formation
• Analysis of pluripotency through Pluritest: Demonstration of stemness using a gene expression array
• Reprogramming factor persistence assays that ensure the loss of exogenous reprogramming factors
• Assessment of iPSC genomic stability by G-banded karyotyping

3. Save on capital expenditure

Outsourcing iPSC reprogramming services can provide significant cost savings. As mentioned earlier, setting up and maintaining an efficient iPSC reprogramming protocol requires a considerable investment in time and resources. Hiring and maintaining staff members to grow, maintain and reprogram cells is an expensive venture which becomes more costly when the cost of reagents and equipment is also added.  By outsourcing to a service that specializes in iPSC generation, researchers can also save money from not having wasted reagents and cells on failed attempts to reprogram them. This cost-saving can be particularly beneficial for small to medium-sized enterprises, as well as academic institutions with limited budgets.

4. Saving time

Meeting deadlines is a priority for every business regardless of its size, which is why outsourcing to a service provider that can generate iPSCs is a powerful reason why researchers are turning to Sampled for iPSC reprogramming services. Instead of trying and failing to reprogram cells over and over again, which inevitably pushes back deadlines, outsourcing gives researchers peace of mind that their iPSCs will arrive on time and is exactly what they need to conduct their investigations. Moreover, outsourcing to a service provider allows researchers to scale up their research without worrying about expanding their lab space, head count or costs associated with purchasing more resources.

In summary, outsourcing iPSC reprogramming services from cells to iPSCs could offer numerous advantages compared to reprogramming iPSCs in house. It saves time, provides access to highly-characterized human iPSCs, produces cells of higher quality, and can provide significant cost savings. Consequently, more organizations and researchers are turning to iPSC generation services to take advantage of these benefits.

To learn more about how our iPSC reprogramming services work, talk to an expert today

The NINDS biobank stem cell collection currently contains 411 subjects, 169 of which have iPSC and 270 of which have fibroblasts and represents several neurodegenerative disorders, including (but not limited to):

• Alzheimer’s disease
• Amyotrophic Lateral Sclerosis
• Frontotemporal Degeneration
• Huntington’s disease
• Parkinson’s disease

If you would like to browse and order cell lines, first go to: https://stemcells.nindsgenetics.org/ and follow the instructions below:

Creating a signed Material Transfer Agreement (MTA)

A key requirement for ordering lines is ensuring a signed MTA is in place.

The MTA forms can be found in the Documents section of the catalog application as shown below.

Selecting cell lines

1. Once you have downloaded the correct MTA form, select the Subjects tab and choose subjects of interest using the filters to narrow by disease, etc, if necessary

Locate the desired cell lines in the subject’s detail view, clicking anywhere within the row will bring up the cell line’s details.

If the subject has cell lines, such as iPSCs and Fibroblasts, they will be displayed in the subject’s detail view. These are found at the bottom of the Full Record tab, and also the Cell Lines tab. Click the Add to Cart button for the desired lines.

Purchasing cell lines

2. Once you have selected the cell line, scroll down and Click Add to Cart.

3. You can then click on the up and down arrows to specify how many vials you would like to purchase (the maximum is 10):

4. Go to the cart and click Checkout.

5. Enter the required fields and upload the required documents, such as signed MTAs (found in the Documents section) and Statement of research intent (SRI).

6. Once all relevant information has been filled in, Click Submit Order 

7. You will receive an e-mail summarizing your order and requesting e-mail address verification.

Here is an example of the e-mail you will receive. Click the link to verify your email address and your order should be all set.

Cell and Data Repository infographic

Download our infographic

View further details on our catalog of the NINDS Human Cell and Data Repository, including a breakdown of gender, race and clinical affected status of cell and disease types.

View infographic

If you need any help ordering from the NINDS website, you can contact us at NINDS@sampled.com and one of our experts will be happy to help.

You can also view our other cellular services here.

Induced pluripotent stem cells (iPSCs) derived from human somatic cells hold massive potential for research into disease-modelling, drug toxicity testing and in regenerative medicine. Thus, as the use of patient-derived stem cells has become more frequently adopted, having a workflow to monitor each cell line is critical. For these reasons, it is imperative that they pass a series of quality control measures to promote reproducibility across experiments and between labs.

To consider these needs, a multistep workflow to assess newly generated iPSCs is vital to test four critical benchmarks:

1. Viability and sterility of cells
2. Genetic abnormalities
3. Stemness
4. Pluripotency (ability to differentiate into the three germ layers)

Testing the sterility

Before using any type of cell line for research it is necessary to ensure that the cells in question are viable and are not contaminated. The biggest entry of species such as Mycoplasma into cell lines are from non-sterilizable cell culture reagents and laboratory personnel. M. orale, M. fermentans, and M. hominis account for more than half of all mycoplasma infections in cell cultures and are found in oropharyngeal tracts¹. Two other species that are known to be prevalent in laboratories are M. arginini and A. laidlawii which can be found in Fetal Bovine Serum and swine-derived Trypsin solutions. Unfortunately, mycoplasma implies resistance against penicillin and can pass 0.2μm sterility filters². This makes it extremely difficult to eradicate mycoplasma infections, furthermore numerous research laboratories do not test for it on a regular basis.  At Sampled however, all iPSC lines are tested for mycoplasma.

G-banded karyotyping and testing for genetic abnormalities

Once the cells have been tested for sterility, it is then necessary to test for genetic abnormalities that could have been introduced into the cells’ DNA during the reprogramming process or induced through culture, giving certain cells a growth advantage. Advances in the field have led to “foot-print-free” reprogramming methods that do not result in reprogramming factor integration, but these new methods do not preclude culture induced genetic mutations. Regardless of which method your iPSCs have been derived, if the cells’ DNA has been compromised, the cells may not be fully functional or perform inadequately.

Furthermore, long-term iPSCs culture can result in chromosomal abnormalities, modification of gene expression, as well as increases the risk of the iPSCs becoming tumorigenic. Since genomic alterations present potential risks in downstream applications, it is important to monitor the genomic integrity of these iPSCs lines especially in iPSCs intended for therapeutic use. It is for these reasons that iPSC karyotype analysis is necessary not just at the start, but throughout your investigation.

Measuring stemness

Immunostaining alone is inefficient at determining stemness since it is difficult to quantify immunostaining results over an entire culture.  At Sampled, we quantitate a large range of stemness markers using Pluritest. This assay uses a the Affymetrix PrimeView Global Gene Expression Array to confirm the undifferentiated state of human pluripotent stem cells.

Fluorescence activated cell sorting analysis (FACS) is another useful method that is commonly employed to measure stemness by measuring expression of stemness markers quantitatively over a large population. FACS relies on the binding of highly specific antibodies tagged with fluorescent dyes to cells expressing stemness markers such as Oct4, Tra-1-60, etc.

Measuring differentiation potential

Lastly, the iPSC’s ability to differentiate into the three germ layers (Ectoderm, Endoderm and Mesoderm layers) ought to be measured before they can be utilized by investigations. In order to test this, the cells need to first be differentiated into Embryoid bodies (EBs). Once the cells have undergone differentiation, the cells are then harvested for gene expression analysis by qPCR and analyzed to ensure that genes specific to all 3 germ layers are expressed.

Is there an easier way to validate your Induced Pluripotent Stem Cells?

It may be daunting for a small or scaling biotech to conduct the tests outlined above before they even begin to start working with these cells. At Sampled, we offer robust quality control services, that can be bundled as part of a package or as bespoke services that includes:

• Sterility testing of all lines includes mycoplasma testing
• Identity verification using the SNPTrace Panel
• Analysis of stemness by FACS
• Demonstration of stemness by Pluritest
• Demonstration of differentiation potential by hPSC scorecard
• Assessment of genomic stability by G-banded karyotyping
• Demonstration of loss of Sendai virus after reprogramming by qPCR

Our highly trained scientists perform these tests to the highest standard and could save your lab time and resources so that you can focus on the research that matters to you the most. If you would like to speak to one of our experts today about validating your stem cells, contact us today.

References

1. Pamies, D., Bal-Price, A., Simeonov, A., Tagle, D., Allen, D., Gerhold, D., Yin, D., Pistollato, F., Inutsuka, T., Sullivan, K. and Stacey, G., 2017. Good cell culture practice for stem cells and stem-cell-derived models. Alternatives to Animal Experimentation: ALTEX, 34(1), pp.95-132.
2. Bruchmüller, I., Pirkl, E., Herrmann, R., Stoermer, M., Eichler, H., Klüter, H. and Bugert, P., 2006. Introduction of a validation concept for a PCR-based Mycoplasma detection assay. Cytotherapy, 8(1), pp.62-69.