Automated, high-throughput derivation, characterization and differentiation of induced pluripotent stem cells

Abstract

Induced pluripotent stem cells (iPSCs) are an essential tool for modeling how causal genetic variants impact cellular function in disease, as well as an emerging source of tissue for regenerative medicine. The preparation of somatic cells, their reprogramming and the subsequent verification of iPSC pluripotency are laborious, manual processes limiting the scale and reproducibility of this technology. Here we describe a modular, robotic platform for iPSC reprogramming enabling automated, high-throughput conversion of skin biopsies into iPSCs and differentiated cells with minimal manual intervention. We demonstrate that automated reprogramming and the pooled selection of polyclonal pluripotent cells results in high-quality, stable iPSCs. These lines display less line-to-line variation than either manually produced lines or lines produced through automation followed by single-colony subcloning. The robotic platform we describe will enable the application of iPSCs to population-scale biomedical problems including the study of complex genetic diseases and the development of personalized medicines.

Integrating cardiomyocytes from human pluripotent stem cells in safety pharmacology: has the time come?

“…The reproducibility of the results in terms of the absolute values of parameters measured may indeed be due to limited standardization of experimental conditions: few studies carry out head‐to‐head comparisons of cells from different hiPSC lines derived under identical conditions. Fully automated, high‐throughput procedures to reprogramme hiPSC from somatic cells have been developed (Paull et al , 2015) with the aim of limiting variability. Pharmacogenomic analysis of these cell populations may unveil the key sources of variability.…”

Good Cell Culture Practice for stem cells and stem-cell-derived models

“…An increasing number of facilities now successfully employs automated platforms capable of feeding, passaging, cryopreserving, and/or selecting colonies (Paull et al, 2015). Automation removes some of the variability inherent to manual culture and enables the maintenance of hundreds of clones in parallel.…”

Human Induced Pluripotent Stem Cells as a Platform for Personalized and Precision Cardiovascular Medicine

“…Beyond the investigation of hiPSC reprogramming mechanisms, clinically relevant advances have been achieved by the development of automated, high-throughput derivation and characterization protocols for hiPSCs (215,245), use of chemically defined media (24), and substitution of retroviral and lentiviral transgene overexpression vectors by integration-free systems that circumvent the risk of spontaneous tumor formation due to insertional mutagenesis. Such methods include the use of excisable viral vectors, nonintegrating viruses (250,251), and even virus-independent approaches such as plasmid, episomal, DNA, protein, or mRNA mediated transgene overexpression (114,293,324).…”

“…In parallel to the evolution of cardiac differentiation protocols in suspension bioreactors as mentioned above (66,132,235,252), automated liquid handling platforms (e.g., Tecan Freedom Evo 100, Sartotious CompacT SelecT, and Beckman Coulter Biomek FXP) for robotic scale-up of mouse and human pluripotent stem cells and their differentiated cardiac derivatives have been evaluated and demonstrated to improve the consistency and quality of cell cultures (105,139,215,267).…”