Neural tissue-spheres: A microexplant culture method for propagation of precursor cells from the rat forebrain subventricular zone

2007 ◽  
Vol 165 (1) ◽  
pp. 55-63 ◽  
Author(s):  
Rikke K. Andersen ◽  
Mathias Johansen ◽  
Morten Blaabjerg ◽  
Jens Zimmer ◽  
Morten Meyer
1999 ◽  
Vol 81 (1) ◽  
pp. 95-102 ◽  
Author(s):  
R. R. Stewart ◽  
T. Zigova ◽  
M. B. Luskin

Stewart, R. R., T. Zigova, and M. B. Luskin. Potassium currents in precursor cells isolated from the anterior subventricular zone of the neonatal rat forebrain. J. Neurophysiol. 81: 95–102, 1999. The progenitor cells from the anterior part of the neonatal subventricular zone, the SVZa, are unusual in that, although they undergo division, they have a neuronal phenotype. To characterize the electrophysiological properties of the SVZa precursor cells, recordings were made of potassium and sodium currents from SVZa cells that were removed from postnatal day 0–1 rats and cultured for 1 day. The properties of the delayed rectifier and A-type potassium currents were described by classical Hodgkin and Huxley analyses of activation and inactivation. In addition, cells were assessed under current clamp for their ability to generate action potentials. The A-type potassium current ( I K(A)) was completely inactivated at a holding potential of −50 mV. The remaining potassium current resembled the delayed rectifier current ( I K(DR)) in that it was blocked by tetraethylammonium (TEA; IC50 4.1 mM) and activated and inactivated slowly compared with I K(A). The conductance-voltage ( G- V) curve revealed that G increased continuously from 0.2 nS at −40 mV to a peak of 2.6 nS at +10 or +20 mV, and then decreased for voltages above +30 mV. Activation time constants were largest at −40 mV (∼11 ms) and smallest at 100 mV (∼1.5 ms). The properties of I K(A) were studied in the presence of 20 mM TEA, to block I K(DR), and from a holding potential of −15 mV, to inactivate both I K(DR) and I K(A). I K(A) was then allowed to recover from inactivation to negative potentials during 200- to 800-ms pulses. Recovery from inactivation was fastest at −130 mV (∼21 ms) and slowest at −90 mV (∼135 ms). Inactivation was voltage independent from −60 to +60 mV with a time constant of ∼15 ms. At steady state, I K(A) was half inactivated at −90 mV. G K(A) increased from 0.2 nS at −60 mV to a peak of 2.4 nS at +40 mV. Finally, the activation time constants ranged from ∼1.9 ms at −50 mV to 0.7 ms at +60 mV. The properties of I K(A) resembled those of I K(A) found in differentiating cerebellar granule neurons. Most SVZa cells had sodium currents (28/32 cells). However, in current clamp 11 of 12 cells were incapable of generating action potentials from voltages of −30 to −100 mV, suggesting that the available current densities were too low to support excitability.


2008 ◽  
Vol 22 (S1) ◽  
Author(s):  
Ty William Abel ◽  
Cara Clark ◽  
Brian Bierie ◽  
Anna Chytil ◽  
Mary Aakre ◽  
...  

Stroke ◽  
2013 ◽  
Vol 44 (3) ◽  
pp. 753-758 ◽  
Author(s):  
Fernando F. Gonzalez ◽  
Amara Larpthaveesarp ◽  
Patrick McQuillen ◽  
Nikita Derugin ◽  
Michael Wendland ◽  
...  

Development ◽  
2001 ◽  
Vol 128 (5) ◽  
pp. 689-702 ◽  
Author(s):  
C.B. Chambers ◽  
Y. Peng ◽  
H. Nguyen ◽  
N. Gaiano ◽  
G. Fishell ◽  
...  

The olfactory bulb, neocortex and archicortex arise from a common pool of progenitors in the dorsal telencephalon. We studied the consequences of supplying excess Notch1 signal in vivo on the cellular and regional destinies of telencephalic precursors using bicistronic replication defective retroviruses. After ventricular injections mid-neurogenesis (E14.5), activated Notch1 retrovirus markedly inhibited the generation of neurons from telencephalic precursors, delayed the emergence of cells from the subventricular zone (SVZ), and produced an augmentation of glial progeny in the neo- and archicortex. However, activated Notch1 had a distinct effect on the progenitors of the olfactory bulb, markedly reducing the numbers of cells of any type that migrated there. To elucidate the mechanism of the cell fate changes elicited by Notch1 signals in the cortical regions, short- and long-term cultures of E14.5 telencephalic progenitors were examined. These studies reveal that activated Notch1 elicits a cessation of proliferation that coincides with an inhibition of the generation of neurons. Later, during gliogenesis, activated Notch1 triggers a rapid cellular proliferation with a significant increase in the generation of cells expressing GFAP. To examine the generation of cells destined for the olfactory bulb, we used stereotaxic injections into the early postnatal anterior subventricular zone (SVZa). We observed that precursors of the olfactory bulb responded to Notch signals by remaining quiescent and failing to give rise to differentiated progeny of any type, unlike cortical precursor cells, which generated glia instead of neurons. These data show that forebrain precursors vary in their response to Notch signals according to spatial and temporal cues, and that Notch signals influence the composition of forebrain regions by modulating the rate of proliferation of neural precursor cells.


2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Angela Maria Cozzolino ◽  
Valeria Noce ◽  
Cecilia Battistelli ◽  
Alessandra Marchetti ◽  
Germana Grassi ◽  
...  

In many cell types, several cellular processes, such as differentiation of stem/precursor cells, maintenance of differentiated phenotype, motility, adhesion, growth, and survival, strictly depend on the stiffness of extracellular matrix that,in vivo, characterizes their correspondent organ and tissue. In the liver, the stromal rigidity is essential to obtain the correct organ physiology whereas any alteration causes liver cell dysfunctions. The rigidity of the substrate is an element no longer negligible for the cultivation of several cell types, so that many data so far obtained, where cells have been cultured on plastic, could be revised. Regarding liver cells, standard culture conditions lead to the dedifferentiation of primary hepatocytes, transdifferentiation of stellate cells into myofibroblasts, and loss of fenestration of sinusoidal endothelium. Furthermore, standard cultivation of liver stem/precursor cells impedes an efficient execution of the epithelial/hepatocyte differentiation program, leading to the expansion of a cell population expressing only partially liver functions and products. Overcoming these limitations is mandatory for any approach of liver tissue engineering. Here we propose cell lines asin vitromodels of liver stem cells and hepatocytes and an innovative culture method that takes into account the substrate stiffness to obtain, respectively, a rapid and efficient differentiation process and the maintenance of the fully differentiated phenotype.


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