Maksymilian Prondzynski, PhD, Instructor, The Department of Cardiology, Boston Children’s Hospital and Harvard Medical School
The Nobel prize winning discovery of human induced pluripotent stem cells (hiPSCs) facilitated a new era of disease modeling in vitro. This event inspired clinicians and scientists alike to use those cells for regenerative medicine, or for modeling of genetic diseases, to gain further insights into disease mechanisms on a cellular level. However, experimental studies using hiPSCs can experience high inter- and intra-experimental variability further fueling the reproducibility crisis in life sciences.
In the past 10 years much progress has been made in defining quality criteria (QC) for hiPSCs to achieve genome stability and high pluripotency over prolonged culture duration. These QC parameters include single cell clonal selection, karyotyping, pluripotency assessment and the establishment of master cell banks, especially in the context of CRISPR-Cas9 genome edited hiPSCs. Additionally, Numerous cardiac differentiation protocols have been established using monolayer (ML) adherent or 3D suspension cultures. However, generating large numbers of high quality human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) with high consistency between batches, different cell lines and cryo-storage has remained challenging leading to poor reproducibility of experimental results.
The featured speakers used a stirred suspension bioreactor system to create an optimal environment for cell growth and differentiation. This enabled them to create a unified differentiation protocol that can be applied to a variety of hiPSCs (patient and control lines) with high cardiomyocyte content and reduced batch-to-batch variability. Using a wide array of cardiac disease modeling assays, they then confirmed high reproducibility of cryo-preserved hiPSC-derived cardiomyocytes.
Join this webinar to learn about optimizing the growth and differentiation of human induced pluripotent stem cells (hiPSCs) into cardiomyocytes and enhancing cardiac disease modeling.
Francisco Ramirez, Manager, Application Science, Bio-Techne
Cloning of Chinese hamster ovary (CHO) cells expressing recombinant antibodies is a crucial step in the development of monoclonal antibody (mAb) therapeutics. It is important to identify clones that produce high yield. However, the heterogeneity of CHO cells can give rise to varied IgG production among individual cells. Thus, the process of finding high titer clones typically involves screening of large numbers of clones, making it laborious and time-consuming.
Cold-capture is a technique that allows labelling of antibody producing cells with a fluorescent reagent that binds to the antibody molecules when they are transiently present on the cell membrane before release from secretory vesicles. This labelling could enable cell sorting based on the level of fluorescence signal to select higher productivity cells. In this study, the featured speaker sought to test the ability to enrich high titer IgG CHO cell clones by cold-capture, followed by isolation of single cells with the brightest fluorescent signal using the Namocell Pala Single Cell Dispenser — a fully automated dispenser that enables a fast, easy fluorescence and light scatter-based single cell isolation. The speaker will show that their method allowed the identification of clones with an average 3x higher IgG production from the cold-capture labelled cells compared to non-cold-captured cells.
These results suggest that the combination of cold-capture and single cell isolation could be an efficient and economical solution for improving cell line development in recombinant antibody production.
Watch this webinar to learn more about the potential of cold-capture-facilitated single cell isolation and cloning to improve the efficiency in identifying high yield clones.
Mason Ouren, R&D Scientist, Luna Genetics