Scientific Publications Featuring
Accutase & Accumax
The selection of studies featured in the articles below show the use of Accutase and Accumax in place of trypsin in the culture of stem cells and other cell types.
Efficient Propagation of Single Cells
Accutase-Dissociated Human Embryonic Stem Cells
hESCs hold great promise for cell-based therapies and drug screening applications. However, growing and processing large quantities of undifferentiated hESCs is a challenging task...This study demonstrates that hESCs can be passaged as single cells using Accutase, a formulated mixture of digestive enzymes. In contrast to trypsin treatment, Accutase treatment does not significantly affect the viability and proliferation rate of hESC dissociation into single cells...Compared to collagenase-passaged hESCs, Accutase-treated cultures contained a larger proportion of undifferentiated (Oct4- positive) cells. Additionally, Accutase-passaged undifferentiated hESCs could be grown as monolayers without the need for monitoring and/or selection for quality hESC colonies. Mol. Reprod. Dev. 2007
Rapid Generation of Functional Dopaminergic Neurons From Human Induced Pluripotent Stem Cells Through a Single-Step Procedure Using Cell Lineage Transcription Factors
Current protocols for in vitro differentiation of human induced pluripotent stem cells (hiPSCs) to generate dopamine (DA) neurons are laborious and time-expensive. In order to accelerate the overall process, we have established a fast protocol....This one-step protocol holds important implications for in vitro disease modeling and is particularly amenable for exploitation in high-throughput screening protocols. Stem Cells Translational Medicine 2013;2:473-479
High Efficacy of Clonal Growth and Expansion of Adult Neural Stem Cells
Neural stem cells (NSCs) from the adult central nervous system are currently being investigated for their potential use in autologous cell replacement strategies. High expansion rates of NSCs in culture are crucial for the generation of a sufficient amount of cells needed for transplantation....In addition, increased cell survival was obtained when Accutase, instead of trypsin, was used for enzymatic dissociation of NSC cultures. This work provides an important step toward the development of standardized protocols for highly efficient in vitro expansion of NSCs from the adult central nervous system to move more closely to the clinical use of NSCs. Lab Invest 2003, 83:949–962
Automated Large-Scale Culture and Medium-Throughput Chemical Screen for Modulators of Proliferation and Viability of Human Induced Pluripotent Stem Cell– Derived Neuroepithelial-like Stem Cells
The aim of this study was to demonstrate proof-of-concept feasibility for the use of human neural stem cells (NSCs) for highthroughput screening (HTS) applications. For this study, an adherent human induced pluripotent stem (iPS) cell–derived long-term, self-renewing, neuroepithelial-like stem (lt-NES) cell line was selected as a representative NSC...Follow-up studies confirmed a dosedependent effect of one of the hit compounds, which was a Cdk-2 modulator. This approach could be further developed as part of a strategy to screen compounds to either improve the procedures for the in vitro expansion of neural stem cells or to potentially modulate endogenous neural stem cell behavior in the diseased nervous system. J Biomol Screen 2013 18: 258
Quantification of cell surface proteins
with bispecific antibodies
Flow cytometry is an established method for fast and accurate quantitation of cellular protein levels and requires fluorescently labeled antibodies as well as calibration standards. A critical step for quantitation remains the production of suitable detection antibodies with a precisely defined ratio of antigen-binding sites to fluorophores...A panel of tumor cell lines was assessed under different culture conditions for absolute receptor expression levels of the indicated antigens and the data were set in relation to mRNA, gene count and immunoblot data. We could reproducibly quantify these receptors, omit the otherwise required normalization step and demonstrate the superiority of a 1 1 1 bispecific antibody. The same antibodies were also used to quantify the number of proteins in intracellular vesicles in confocal microscopy. The antibodies can be stored like regular antibodies and can be coupled with different digoxigeninlabeled fluorophores which makes them excellent tools for FACS and imaging-based experiments.