Dissociating Tissues into Cells and the Development of Hydra from Aggregated Cells

Flick, K.; Bode, Hans R.

doi:10.1007/978-1-4757-0596-6_33

Cited by 12 publications

(7 citation statements)

References 4 publications

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“…The same region observed under bright field shows that the fluorescence is not located within nematocytes ( Figure 2D ). Analysis of single viable or fixed cells obtained from dissociation and maceration procedures [35] , [36] , respectively, confirmed the intracellular localization of QRs ( Figure 3 ). The use of both procedures to examine single cell suspension led to detection of membrane labelling only in living dissociated cells, and not in fixed cells, either due to experimental artefacts, or to a fast effect of the chemicals used for fixation, perhaps pH dependent, on membrane trafficking.…”

Section: Resultsmentioning

confidence: 55%

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Fluorescent Nanocrystals Reveal Regulated Portals of Entry into and Between the Cells of Hydra

et al. 2009

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Initially viewed as innovative carriers for biomedical applications, with unique photophysical properties and great versatility to be decorated at their surface with suitable molecules, nanoparticles can also play active roles in mediating biological effects, suggesting the need to deeply investigate the mechanisms underlying cell-nanoparticle interaction and to identify the molecular players. Here we show that the cell uptake of fluorescent CdSe/CdS quantum rods (QRs) by Hydra vulgaris, a simple model organism at the base of metazoan evolution, can be tuned by modifying nanoparticle surface charge. At acidic pH, amino-PEG coated QRs, showing positive surface charge, are actively internalized by tentacle and body ectodermal cells, while negatively charged nanoparticles are not uptaken. In order to identify the molecular factors underlying QR uptake at acidic pH, we provide functional evidence of annexins involvement and explain the QR uptake as the combined result of QR positive charge and annexin membrane insertion. Moreover, tracking QR labelled cells during development and regeneration allowed us to uncover novel intercellular trafficking and cell dynamics underlying the remarkable plasticity of this ancient organism.

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Section: Resultsmentioning

confidence: 55%

“…Hydra polyps were dissociated according to Flick and Bode [35] with minor modifications. This procedure allows to dissociate Hydra tissue into a suspension of viable single cells.…”

Section: Methodsmentioning

confidence: 99%

Fluorescent Nanocrystals Reveal Regulated Portals of Entry into and Between the Cells of Hydra

et al. 2009

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“…The resulting body columns were placed for $2.5 min in ice-cold HM solution, with the pH adjusted to 2.5 using 2 M HCl, then transferred to Dissociation Medium (DM) composed of 3.6 mM KCl (Research Products International, Mt. Prospect, IL), 6 mM CaCl 2 (Spectrum), 1.2 mM MgSO 4 (Fisher Bioreagents), 6 mM sodium citrate (LabChem, Inc., Zelienople, PA), 6 mM sodium pyruvate (Alfa Aesar, Ward Hill, MA), 6 mM glucose (Sigma, St. Louis, MO), 12.5 mM TES (Sigma), 50 mg/mL rifampicin (Calbiochem, San Diego, CA), at pH 6.9 at room temperature (RT), following (30). The dishes containing the body columns in DM were sealed with tape and swirled to promote separation of tissues.…”

Section: Tissue Separationmentioning

confidence: 99%

Physical Mechanisms Driving Cell Sorting in Hydra

Cochet‐Escartin

Locke

Shi

et al. 2017

Biophysical Journal

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Cell sorting, whereby a heterogeneous cell mixture organizes into distinct tissues, is a fundamental patterning process in development. Hydra is a powerful model system for carrying out studies of cell sorting in three dimensions, because of its unique ability to regenerate after complete dissociation into individual cells. The physicists Alfred Gierer and Hans Meinhardt recognized Hydra's self-organizing properties more than 40 years ago. However, what drives cell sorting during regeneration of Hydra from cell aggregates is still debated. Differential motility and differential adhesion have been proposed as driving mechanisms, but the available experimental data are insufficient to distinguish between these two. Here, we answer this longstanding question by using transgenic Hydra expressing fluorescent proteins and a multiscale experimental and numerical approach. By quantifying the kinematics of single cell and whole aggregate behaviors, we show that no differences in cell motility exist among cell types and that sorting dynamics follow a power law with an exponent of ∼0.5. Additionally, we measure the physical properties of separated tissues and quantify their viscosities and surface tensions. Based on our experimental results and numerical simulations, we conclude that tissue interfacial tensions are sufficient to explain cell sorting in aggregates of Hydra cells. Furthermore, we demonstrate that the aggregate's geometry during sorting is key to understanding the sorting dynamics and explains the exponent of the power law behavior. Our results answer the long standing question of the physical mechanisms driving cell sorting in Hydra cell aggregates. In addition, they demonstrate how powerful this organism is for biophysical studies of self-organization and pattern formation.

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“…Three solutions were prepared: 1% solution of procaine-HCl (Wako Pure Chemicals, Tokyo) in distilled water, hyper-osmotic (70 mosM) salt solution for tissue dissociation and cell reaggregation (DM solution; Flick and Bode, 1983), and Hydra culture solution described above. Equal volumes of the three solutions were mixed and adjusted to pH.…”

Section: Separation Of Ectodermal and Endodermal Tissue Layersmentioning

confidence: 99%

Motility of endodermal epithelial cells plays a major role in reorganizing the two epithelial layers in Hydra

Takaku

Hariyama

Fujisawa

2005

Mechanisms of Development

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Cell-cell interactions and cell rearrangements play important roles during development. Aggregates of Hydra cells reorganize into the two epithelial layers and subsequently form a normal animal. Examination of the formation of the two layers under various situations, indicates that the motility of endodermal epithelial cells, but not the differential adhesive forces of the two types of epithelial cells, plays the critical role in setting up the two epithelial layers. (1) When aggregates of ectodermal cells and of endodermal cells were placed in direct contact, the endodermal cells migrated into the interior of the ectodermal aggregate. This process was completely inhibited by cytochalasin B although initial firm attachment between the two aggregates was not blocked. (2) A single endodermal epithelial cell placed in contact with an ectodermal aggregate, actively extended pseudopod-like structures and migrated toward the center of the ectodermal aggregate. In contrast, an ectodermal epithelial cell remained in contact with an endodermal aggregate and never exhibited migratory behavior. Cytochalasin treatment of only endodermal epithelial cells abolished the migration. (3) One to 4 endodermal epithelial cells and/or ectodermal epithelial cells were placed in contact with one another forming up to 4-cell aggregates. Endodermal epithelial cells exhibited high motility that can be attributed to the migratory movement described above. Finally, formation of actin bundles, as visualized with rhodamine-phalloidin, was always correlated with pseudopod formation in endodermal epithelial cells during early and mid stages of aggregate formation.

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Dissociating Tissues into Cells and the Development of Hydra from Aggregated Cells

Cited by 12 publications

References 4 publications

Fluorescent Nanocrystals Reveal Regulated Portals of Entry into and Between the Cells of Hydra

Fluorescent Nanocrystals Reveal Regulated Portals of Entry into and Between the Cells of Hydra

Physical Mechanisms Driving Cell Sorting in Hydra

Motility of endodermal epithelial cells plays a major role in reorganizing the two epithelial layers in Hydra

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