Induced pluripotent stem (iPS) cells are one of the most valuable tools in biomedical research. These special cells originate from regular somatic cells, like those comprising skin. By ectopically expressing key transcription factors, the cells are reprogrammed to a pluripotent state resembling embryonic stem cells. Researchers can then propagate the iPS cells indefinitely and differentiate them when needed into virtually any cell type. This versatility has made iPS cells invaluable to a variety of scientific fields—including regenerative and disease medicine—as researchers can more easily obtain cell types that resemble primary tissues.

One of the most widely-used applications of iPS cells is disease modeling. Using the powerful gene-editing tool, CRISPR, researchers can introduce specific mutations to healthy cells to recapitulate diseases. Alternatively, one can revert the genotype of cells taken from individuals with a disease to wild type. For instance, Dr. Ellerby at the Buck Institute uses iPS cells-derived neuronal cells as model systems to mimic neurological diseases such as Huntington’s, Alzheimer’s, and Parkinson’s (learn more here).

An additional benefit of iPS cells is that they can be generated from somatic cells sourced from patents, enabling the generation of models and isogenic cell lines with patient-specific genetic backgrounds. With this unique capability, iPS cell technology is shaping the future of personalized medicine.

Although iPS cells offer researchers great opportunities for understanding diseases, genetically altering them while maintaining a high standard of quality is not easy. Difficulties may arise in keeping the cells alive, undifferentiated, and genetically stable. Moreover, the CRISPR editing workflow requires high-quality reagents, optimized transfection procedures, and stringent quality control. If clones are needed, then the workflow includes extra hands-on time and constant monitoring over weeks. Any of these challenges can serve as a significant barrier to capitalizing on the benefits of iPS cells.

 

Keeping iPS Cells Pluripotent Can Be Difficult!

Induced pluripotent stem (iPS) cells are one of the most valuable tools in biomedical research. These special cells originate from regular somatic cells, like those comprising skin. By ectopically expressing key transcription factors, the cells are reprogrammed to a pluripotent state resembling embryonic stem cells. Researchers can then propagate the iPS cells indefinitely and differentiate them when needed into virtually any cell type. This versatility has made iPS cells invaluable to a variety of scientific fields—including regenerative and disease medicine—as researchers can more easily obtain cell types that resemble primary tissues.

One of the most widely-used applications of iPS cells is disease modeling. Using the powerful gene-editing tool, CRISPR, researchers can introduce specific mutations to healthy cells to recapitulate diseases. Alternatively, one can revert the genotype of cells taken from individuals with a disease to wild type. For instance, Dr. Ellerby at the Buck Institute uses iPS cells-derived neuronal cells as model systems to mimic neurological diseases such as Huntington’s, Alzheimer’s, and Parkinson’s (learn more here).

An additional benefit of iPS cells is that they can be generated from somatic cells sourced from patents, enabling the generation of models and isogenic cell lines with patient-specific genetic backgrounds. With this unique capability, iPS cell technology is shaping the future of personalized medicine.

Although iPS cells offer researchers great opportunities for understanding diseases, genetically altering them while maintaining a high standard of quality is not easy. Difficulties may arise in keeping the cells alive, undifferentiated, and genetically stable. Moreover, the CRISPR editing workflow requires high-quality reagents, optimized transfection procedures, and stringent quality control. If clones are needed, then the workflow includes extra hands-on time and constant monitoring over weeks. Any of these challenges can serve as a significant barrier to capitalizing on the benefits of iPS cells.

 

Keeping iPS Cells Pluripotent Can Be Difficult!

Synthego Solution: CRISPR-Edited iPS Cells

Fortunately, Synthego can help! Leveraging their automation capabilities, Synthego editing workflow includes several steps that ensure high editing efficiencies while maintaining a high standard of quality. As seen in the illustration below, Synthego employs a streamlined workflow that can produce edited cell pools and clones. Edited pools are cell populations that contain both edited and wild type cells. Clones are populations of genetically identical cells that contain the desired edit.

Several steps in the editing process enable Synthego to produce edited iPS cells of the highest quality. Let’s take a closer look:

  1. Guide Synthesis: all guide RNAs are produced synthetically and contain chemical modifications that increase stability and lower immunogenicity.
  2. Transfection: guide RNA and Cas9 nuclease are introduced to cells as ribonucleoproteins (RNPs) using electroporation. Because RNPs cut the DNA quickly but for a limited period of time, the risk of off-target editing is decreased. Electroporation utilizes electrical impulses to introduce RNPs into cells and enables high editing efficiencies in a variety of cell types.
  3. Quality Control: Stringent quality control checks are integral parts of the workflow for both pools and clones. Edited pools are checked for purity and pluripotency, and clones are checked for purity, pluripotency, and genomic stability (karyotyping).
    Synthego’s Quality Control Measures:

     

    Measure Purpose
    PluriTest Evaluates pluripotency
    Sterility test Detects mycoplasma/virus contamination
    KaryoStat Assesses genomic stability/ diploid state
  4. Clonal Isolation: Synthego uses proprietary imaging technology to ensure that each clone originates from a single cell and that selected clones are pluripotent.

Learn how to get high-quality iPS cells with your desired edit, guaranteed.

Overcome Barriers of Gene Editing in iPS Cells

Induced pluripotent stem (iPS) cells offer unprecedented access to cell types that resemble primary tissues. Coupled with the power of CRISPR, researchers can now utilize these cells to generate highly specific genetically-engineered models.

Despite their capabilities, working with iPS cells is tricky, and making edits while keeping them happy is even harder. Moreover, isolating edited clones is a tedious process that can take months of valuable time. Fortunately, Synthego can help. With their rigorous editing workflow, Synthego guarantees delivery of your desired edit in fully pluripotent, stable iPS cells.