This protocol details a crucial method for accurately characterizing and purifying human mesenchymal stem cells on a single platform, enabling efficient downstream applications like stem cell transplantation. It is particularly significant for clinical labs enhancing molecular and cytogenetic assays through precise single cell sorting of engineered or natural stem cells. The commonly used methods of isolation of purified stem cell populations refer to techniques such as immunomagnetic separation, density gradient, or microfluidic cell sorting.
A technique like index sorting allows it to be linked to single cell RNA profiling that can extract transcriptomic information of each sorted cell. The key challenges involve maintaining sample quality, optimization of cell culture conditions, including cell density and viability during and after sorting is required. Additionally, choosing suitable nozzle sizes for the instrument and ensuring timely access to the sorters would also be considered.
This protocol illustrates a precise staining panel for immunophenotyping and sorting the desired cell population. It ensures standardized sample quality, optimal cell viability, density for cell population sorting and define sorting experimental parameters on the FACSDiva software. This method enables a straightforward selection and immunophenotyping of stem cell population expressing specific biomarkers.
This simple platform allows efficient sorting based on the target population aiding and obtaining purified samples for subsequent downstream applications.
