Automated generation of human stem cell clones by Image-Activated Cell Selection (IACS™)

Koch, Philipp; Biegler, Andrea; Pfennig, Annette; Müller, Torsten; Klein-Vehne, Anne; Terstegge, Stefanie; Itskovitz‐Eldor, Joseph; Brüstle, Oliver

doi:10.1038/nmeth809

Cited by 2 publications

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“…Single neural precursors derived from lines H9.2 (passages 25-75) and I3 (passages 20-60) were deposited on gamma-irradiated astrocytes derived from murine ESCs using the CytoClone system (9,27) (Fig. S4).…”

Section: Resultsmentioning

confidence: 99%

A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration

Koch

Opitz²,

Steinbeck³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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An intriguing question in human embryonic stem cell (hESC) biology is whether these pluripotent cells can give rise to stably expandable somatic stem cells, which are still amenable to extrinsic fate instruction. Here, we present a pure population of long-term self-renewing rosette-type hESC-derived neural stem cells (lt-hESNSCs), which exhibit extensive self-renewal, clonogenicity, and stable neurogenesis. Although lt-hESNSCs show a restricted expression of regional transcription factors, they retain responsiveness to instructive cues promoting the induction of distinct subpopulations, such as ventral midbrain and spinal cord fates. Using lt-hESNSCs as a donor source for neural transplantation, we provide direct evidence that hESC-derived neurons can establish synaptic connectivity with the mammalian nervous system. Combining long-term stability, maintenance of rosette-properties and phenotypic plasticity, lt-hESNSCs may serve as useful tool to study mechanisms of human NSC self-renewal, lineage segregation, and functional in vivo integration.human embryonic stem cells ͉ neural differentiation ͉ regionalization ͉ synapse formation

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Section: Resultsmentioning

confidence: 99%

A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration

Koch

Opitz²,

Steinbeck³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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450

504

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High‐resolution 3‐D imaging of living cells in suspension using confocal axial tomography

Renaud

Viña²,

et al. 2008

Biotechnology Journal

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Conventional flow cytometry (FC) methods report optical signals integrated from individual cells at throughput rates as high as thousands of cells per second. This is further combined with the powerful utility to subsequently sort and/or recover the cells of interest. However, these methods cannot extract spatial information. This limitation has prompted efforts by some commercial manufacturers to produce state-of-the-art commercial flow cytometry systems allowing fluorescence images to be recorded by an imaging detector. Nonetheless, there remains an immediate and growing need for technologies facilitating spatial analysis of fluorescent signals from cells maintained in flow suspension. Here, we report a novel methodological approach to this problem that combines micro-fluidic flow, and microelectrode dielectric-field control to manipulate, immobilize and image individual cells in suspension. The method also offers unique possibilities for imaging studies on cells in suspension. In particular, we report the system's immediate utility for confocal "axial tomography" using micro-rotation imaging and show that it greatly enhances 3-D optical resolution compared with conventional light reconstruction (deconvolution) image data treatment. That the method we present here is relatively rapid and lends itself to full automation suggests its eventual utility for 3-D imaging cytometry.

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Automated generation of human stem cell clones by Image-Activated Cell Selection (IACS™)

Cited by 2 publications

References 0 publications

A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration

A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration

High‐resolution 3‐D imaging of living cells in suspension using confocal axial tomography

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