Re: Vannoni presenta il suo Metodo Stamina
Inviato da Ste_79 il 5/10/2014 21:40:38
Citazione:
Grazie a te che hai permesso di sviscerare un po' di più l'argomento.
Aggiornerò questo post ogni tanto. Quando vuoi dacci un occhio le tue conoscenze mi potrebbero tornare utili.
Citazione:
L'unica cosa che ho trovato è presente nella richiesta di brevetto ed è la seguente:
a) Differentiation into Neuroblasts/Neurons in Suspension
To the mesenchymal stem cells suspended in physiological solution 6 μl/ml of neuronal differentiation solution are added.
The solution obtained is delicately resuspended and maintained at 37° C. for a period comprised between 20 minutes and 2 hours, preferably between 40 minutes and 1 hour and 30 minutes, in function of the maturation state (neuroblasts-neurons) desired.
At two hours mature neurons with dendrites and axons completely formed and functional are present (see FIGS. 1 and 2 above).
The solution for neuronal differentiation is composed thusly:
10 ml of 98% ethanol
10 mg of retinoic acid.
The relative quantities indicated above are to be considered exclusively as a practical indication for obtaining a differentiation solution having a retinoic acid concentration of 3×10−3 M.
The neuronal differentiation solution is agitated to dissolve the retinoic acid and maintained refrigerated at 4° C.
b) Differentiation into Neuroblasts/Neurons in Adhesion
Six microlitres per millilitre (6 μl/ml) of the neuronal differentiation solution prepared just before use (maximum 1 hour) and stored at +4° C. in the dark is added to the mesenchymal stem cells adherent to the wall of a culture flask.
The flask is placed in an incubator for a period comprised between 20 minutes and 2 hours, preferably between 40 minutes and 1 hour and 30 minutes, in function of the maturation state (neuroblasts-neurons) desired.
At two hours mature neurons are present with completely formed and functional dendrites and axons, the morphology of which can be appreciated (see FIGS. 3 and 4) and on which it is possible to conduct immunohistochemical and electrophysical tests.
The solution for neuronal differentiation is composed thusly:
10 ml of 98% ethanol
10 mg of retinoic acid.
The relative quantities indicated above are to be considered exclusively a practical indication for obtaining a differentiation solution having a retinoic acid concentration of 3×10−3 M.
Brevetto
ovviamente non ti so dire se è sufficiente come spiegazione per un professore, anche se credo fortemente di no, per tornare al discorso più sopra.
Interessante.
Seguirò gli sviluppi quando possibile.
Grazie a te che hai permesso di sviscerare un po' di più l'argomento.
Aggiornerò questo post ogni tanto. Quando vuoi dacci un occhio le tue conoscenze mi potrebbero tornare utili.
Citazione:
Ok, ma esiste una formula dettagliata ?
Tipo un protocollo da seguire per fare la soluzione ?
L'unica cosa che ho trovato è presente nella richiesta di brevetto ed è la seguente:
a) Differentiation into Neuroblasts/Neurons in Suspension
To the mesenchymal stem cells suspended in physiological solution 6 μl/ml of neuronal differentiation solution are added.
The solution obtained is delicately resuspended and maintained at 37° C. for a period comprised between 20 minutes and 2 hours, preferably between 40 minutes and 1 hour and 30 minutes, in function of the maturation state (neuroblasts-neurons) desired.
At two hours mature neurons with dendrites and axons completely formed and functional are present (see FIGS. 1 and 2 above).
The solution for neuronal differentiation is composed thusly:
10 ml of 98% ethanol
10 mg of retinoic acid.
The relative quantities indicated above are to be considered exclusively as a practical indication for obtaining a differentiation solution having a retinoic acid concentration of 3×10−3 M.
The neuronal differentiation solution is agitated to dissolve the retinoic acid and maintained refrigerated at 4° C.
b) Differentiation into Neuroblasts/Neurons in Adhesion
Six microlitres per millilitre (6 μl/ml) of the neuronal differentiation solution prepared just before use (maximum 1 hour) and stored at +4° C. in the dark is added to the mesenchymal stem cells adherent to the wall of a culture flask.
The flask is placed in an incubator for a period comprised between 20 minutes and 2 hours, preferably between 40 minutes and 1 hour and 30 minutes, in function of the maturation state (neuroblasts-neurons) desired.
At two hours mature neurons are present with completely formed and functional dendrites and axons, the morphology of which can be appreciated (see FIGS. 3 and 4) and on which it is possible to conduct immunohistochemical and electrophysical tests.
The solution for neuronal differentiation is composed thusly:
10 ml of 98% ethanol
10 mg of retinoic acid.
The relative quantities indicated above are to be considered exclusively a practical indication for obtaining a differentiation solution having a retinoic acid concentration of 3×10−3 M.
Brevetto
ovviamente non ti so dire se è sufficiente come spiegazione per un professore, anche se credo fortemente di no, per tornare al discorso più sopra.
Messaggio orinale: https://old.luogocomune.net/site/newbb/viewtopic.php?forum=6&topic_id=7561&post_id=262362