Time-lapse analysis of human embryonic stem cells reveals multiple bottlenecks restricting colony formation and their relief upon culture adaptation. Increased risk of genetic and epigenetic instability in human embryonic stem cells associated with specific culture conditions. Basic fibroblast growth factor support of human embryonic stem cell self-renewal. Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment. Chemically defined conditions for human iPSC derivation and culture. Derivation of human embryonic stem cells in defined conditions. Feeder-independent culture of human embryonic stem cells. Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. A small-molecule cocktail that beats cellular stress. Establishment in culture of pluripotential cells from mouse embryos. Multi-lineage human iPSC-derived platforms for disease modeling and drug discovery. Induced pluripotent stem cell technology: a decade of progress. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Embryonic stem cell lines derived from human blastocysts. Altogether, the relative ease, scalability and robustness of this workflow should boost gene editing in hPSCs and leverage a wide range of applications, including cell line development (e.g., reporter and isogenic cell lines), disease modeling and applications in regenerative medicine. Depending on the proliferation rate of the clone derived from a single cell, this protocol can be completed in 7–14 d and requires experience with aseptic cell culture techniques. The protocol also facilitates clone picking and produces genetically stable clonal cell lines from hPSCs in a safe and cost-efficient fashion. The use of low-pressure microfluidic cell dispensing ensures gentle and rapid dispensing of single cells into 96- and 384-well plates, while the fast-acting CEPT cocktail minimizes cellular stress and maintains cell structure and function immediately after cell dissociation. This advanced strategy promotes the viability and cell fitness of self-renewing stem cells. Here we describe a chemically defined protocol for robust single-cell cloning using microfluidics-based cell sorting in combination with the CEPT small-molecule cocktail. This inefficiency of cell cloning represents a major obstacle for the standardization and streamlining of gene editing in induced pluripotent stem cells for basic and translational research. Single-cell dissociation and the establishment of clonal cell lines have been long-standing challenges. The PBS-MINI MagDrive Bioreactor Base Unit includes a power adapter that is compatible with Type A electrical outlets and 120V or 240V A/C inputs.Human pluripotent stem cells (hPSCs) are inherently sensitive cells. provides an 18-month warranty for the PBS-MINI MagDrive Bioreactor Base Unit, valid from date of shipment. These disposable vessels are animal component-free and meet the requirements for USP Class VI Testing for Plastics and ISO 10993. PBS-MINI 0.1 L and 0.5 L MAG Single-Use Vessels are sold separately in packs of 4. Conveniently control your culture system with a speed dial and digital display, and visualize cells in low-light conditions using built-in LED lights. Ideal for human pluripotent stem cells (hPSCs), the compact, sealed Base Unit can be used inside incubators. The gentle yet efficient mixing provided by the Vertical-Wheel™ impeller enables the expansion of shear-sensitive cells without anti-foaming agents or shear protectants. Reliably and rapidly scale up your 3D cell cultures and suspensions with the PBS-MINI MagDrive Bioreactor Base Unit and MAG Single-Use Vessels. Tissue and Cell Culture Dissociation Reagents.Work at STEMCELL View Current Opportunities >
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