Induced pluripotent and embryonic stem cell research workflow for culture and maintenance

High-quality human pluripotent stem cell (PSC) lines, either embryonic or induced pluripotent stem cells, are the foundation for any successful and reproducible PSC research. Choosing the most supportive cell culture conditions are critical when expanding, passaging, or cryopreserving PSCs, as only high-quality lines show pluripotent characteristics and successfully pass all mandatory characterization assays.

Balanced and carefully optimized media formulations are key features for high-quality PSC maintenance. StemMACS™ iPS-Brew XF, human and StemMACS PSC-Brew XF, human have been specifically designed to provide your cells with the best-quality nutrients to sustain robust growth and maintain a high pluripotent phenotype and differentiation potential.

Pluripotent stem cells cultured in StemMACS PSC-Brew XF display typical colony morphology (A) and high expression of pluripotency markers (B).

StemMACS PSC-Brew XF, human

Ensuring a state-of-the-art PSC culture with stable doubling times, our products provide high expression of pluripotency markers, full pluripotent potential, and a stable karyotype. Our solutions enable hPSCs grown in StemMACS PSC-Brew XF, human the ability to show typical morphology, expression of pluripotency-associated markers and retain the ability to differentiate into cell types that derive from the three embryonic germ layers, even after consecutive passages in culture.

Overview of flexible feeding schedules with StemMACS PSC-Brew, human and StemMACS iPS-Brew XF, human.

Feeding schedule

Moreover, StemMACS PSC-Brew XF, human gives you the possibility of free weekends, allowing more time for what matters. With optimized nutrient supply, stable pH, and FGF-2 IS you can now routinely skip feeding on weekends and rely on an every other day schedule throughout the week to meet your needs. Additional combinations with StemMACS PSC Support XF also provide the opportunity for cutting edge applications, such as gene editing or cell sorting, providing complete support throughout your workflow.

After some days in culture, PSCs will reach confluence triggering the need to split into additional culture plates (passaging). Splitting the PSC culture is a delicate time point and specific techniques and reagents are necessary for maintaining a stable line. Classic passaging techniques, such as the use of trypsin, are not suitable due to its tendency to damage PSC epitopes usually generating single cell suspension, a condition not well tolerated by PSCs.

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Passaging of Pluripotent Stem Cells

Contrastingly to traditional techniques, StemMACS Passaging Solution XF ensures gentle detachment of PSC colonies and dissociation as cell clusters, therefore preserving cell-cell contacts. The solution is designed to minimize manipulation of the culture, eliminating inactivation, dilution, or centrifugation steps. 

PSCs must show a specific characterization for embryonic stem cell (ESC) or induced pluripotent stem cell (iPSC) lines. Importantly, these characteristics must be established during propagation of already established lines in order to detect early signs of spontaneous differentiation. Typical PSC characterization includes assessment of extra- and intracellular marker expression as well as assessment of pluripotent differentiation potential. 

Multicolor flow cytometry analysis of undifferentiated human iPSCs.

Expression of pluripotency-associated markers

PSCs must show high and stable expression of a combination of pluripotency associated markers. To make this assessment easier and faster, we have created a multicolor panel for flow cytometry. By using it, you can benefit from the simultaneous quantification of intracellular and surface markers. Importantly, you will obtain not only qualitative but also quantitative data.

Workflow for pluripotency assessment with StemMACS Trilineage Differentiation Kit, human.

Functional assessment of pluripotency in just seven days

PSCs by definition, need to be able to differentiate into cellular types that derive from the 3 embryonic germ layers (endoderm, mesoderm, ectoderm). This property needs to be routinely assessed, to prove the highest quality of the line. To make this task easier and even more accurate, we have developed a standardized and quantifiable differentiation assay based on lineage-specific complete media that directly supports 2D differentiation into all three germ layers within seven days. 

The StemMACS Trilineage Differentiation Kit, human contains ready-to-use media to ensure reproducibility and minimizes your effort allowing a definitive side-by-side comparison of different cell lines or clones. 

Quantitative flow cytometric analysis of trilineage differentiation using the StemMACS Trilineage Differentiation Kit, human

Multicolor panel for flow cytometry with REAfinity™ Recombinant Antibodies

We have also extended this innovation by quickening analysis capabilities with our multicolor panel for flow cytometry with REAfinity Recombinant Antibodies. The antibody panel now allows for rapid but sensitive, qualitative, and quantitative analysis of hPSC differentiation potential. Consisting of two antibody combinations for each embryonic germ layer (ectoderm, mesoderm, and endoderm) the ability to detect both surface and intracellular markers are now accessible.

StemMACS Cryo‑Brew ensures high recovery and viability after cryopreservation of induced pluripotent stem cells (iPSCs)

Optimal storage and cryopreservation is essential for the stability and accuracy of many cell culture workflows. The importance of trusting such a medium to maintain these characteristics is vital in workflow success.  StemMACS Cryo-Brew is a cryopreservation medium that has been designed for use with xeno- and serum- free culture systems ensuring high recovery, viability, and genetic stability of PSCs.

Get acquainted with our protocols and antibody panels for your workflows in pluripotent stem cell research.

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