Application note by cytena

f.sight | Streamlined Workflow for the Generation of CRISPR-edited Mesenchymal Stem Cell Clones for Regenerative Medicine Applications

Here we see demonstrated an efficient, high-throughput workflow that incorporates cytena’s f.sight for both selection and cloning of CRISPR/Cas9-edited mesenchymal stem cells (MSCs). Here, MSCs underwent gene editing to knockout the RANKL protein to further enhance the therapeutic capacity of MSCs in regenerative medicine. The f.sight’s high sensitivity was able to detect the low fluorescent signal of the edited MSCs and enrich for this population. The high single-cell isolation efficiency (>93%) and gentle isolation of the f.sight enabled the generation of 182 monoclonal cell lines. Of these, 17 were selected for further characterization. Two successfully edited clones that exhibited the capacity for osteogenic differentiation were generated.

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