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Identifying genes


Tissue culture
Renato Dulbecco Scientist
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So, we embarked upon it, we also decided to continue to study the development in a simpler system, and the simpler system was provided by tissue cultures. We're back with the rat again, where I studied previously and there was a culture that was derived from another person and this culture was the culture that developed in many directions. And therefore I closely analysed all the various sub-types that emerged and I saw that there were a few that were really stable and in particular, there were two stable types; one, the cells were truly epithelial, are the cancer cells, the cells from the breast ducts, and the other, however, with the cells that seemed fusiform, not therefore epithelial, that had to be a type... it was difficult to say to what they corresponded, in short. But there were two stable types and then I kept these cultures. And, observing the cultures of these epithelial cells, I noted that when the cells grew in the cultures, on glass plates, the cells always became more numerous, they started to develop what we call domes, therefore they broke away from the substrate and grew forming this dome. And these domes had a structure, a completely different appearance from the other cells, therefore it was clearly a phenomenon of differentiation. I thought to myself that this was interesting, that we could use this to study differentiation; however, the mammary gland was so complex that it was difficult to try and understand everything. We started to study this and we studied the genes involved. And we did this by comparing these two cultures that I was talking about: the one that makes the domes and the other fusiform, so we obtained messengers from two types of cells and then we put one in large quantities and the other in small quantities.

[Q] So both in epithelial and in fusiform?

Yes, we took considerable quantities of the one from the epithelial cells, and the other in small quantities and then we had the small amount absorbed by the large amounts so that what remained were only cells that were expressed in one but not in the other, do you see? Thus, we obtained a series of genes from one and from the other type of cell and using these types of genes of the epithelial cell, we managed to identify several genes that are involved in the formation of these domes.

Perciò ci siamo imbarcati, anche lì abbiamo deciso di continuare a studiare lo sviluppo in un sistema più semplice e il sistema più semplice era dato da culture di tessuti. Siamo di nuovo di ritorno al ratto, dove io avevo studiato prima e c'era una cultura che è stata derivata da un'altra persona e che questa cultura era la cultura che si sviluppava in molte direzioni. E perciò io ho analizzato bene tutti i vari sottotipi che venivano fuori da questo e ho visto che ce n'erano alcuni che erano veramente stabili e specialmente ce n'erano due tipi stabili: uno, le cellule veramente proprio veramente epiteliali, sono le cellule del cancro, le cellule dei dotti mammari. E un l'altro, invece, con le cellule che sembravano fusiformi, non perciò epiteliali, che dovevano essere un tipo... era difficile dire a che cosa corrispondessero. Insomma, ma erano due tipi stabili e allora li ho mantenuti a queste culture. E, osservando le culture di queste cellule epiteliali, notavo che, quando le cellule crescevano nelle culture, su piastre di vetro, diventavano sempre le cellule più numerose, cominciavano a sviluppare quelle che chiamano i 'domes', delle cupole, perciò si distaccavano dal substrato e crescevano formando questa cupola. E queste cupole avevano una struttura, un aspetto completamente diverso dalle altre cellule, perciò era evidentemente un fenomeno di differenziazione. Ho pensato questo è una cosa interessante, possiamo usare questo per studiare la differenziazione, invece la ghiandola mammaria come tale è così complessa che è difficile andare a capire tutti i punti che ci sono; cominciamo a studiare questo e studiamo i geni che sono coinvolti. E questo l'abbiamo fatto paragonando queste due culture che dicevo: quella che faceva i 'domes' e l'altra fusiforme, perciò noi ottenevamo dei messaggeri dai due tipi di cellule e poi mettevamo uno in grande quantità e l'altro in piccola quantità.

[Q] Allora, sia nelle epiteliali, sia nelle fusiformi?

Sì, quello delle cellule epiteliali, prendevamo notevoli quantità e l'altro piccola quantità e poi facevamo assorbire questa piccola quantità da quella grande quantità, per cui quello che rimaneva erano solo cellule che erano espresse in una ma non nell'altra, capisci? Così, abbiamo ottenuto una serie di geni dell'uno e dell'altro tipo di cellula e usando questi tipi di geni della cellula epiteliale, siamo riusciti a identificare parecchi geni che sono coinvolti nella formazione di questi 'domes'.

The Italian biologist Renato Dulbecco (1914-2012) had early success isolating a mutant of the polio virus which was used to create a life-saving vaccine. Later in his career, he initiated the Human Genome Project and was jointly awarded the Nobel Prize in Physiology or Medicine in 1975 for furthering our understanding of cancer caused by viruses.

Listeners: Paola De Paoli Marchetti

Paola De Paoli Marchetti is a science journalist who graduated with an honours degree in foreign languages and literature from the University Ca’Foscari, Venice. She has been a science journalist since the 1960s and has been on the staff of the newspaper Il Sole 24 Ore since 1970. She was elected president of UGIS (Italian Association of Science Journalists) in 1984. She has been a Member of the Board of EUSJA (European Union of Science Journalists’ Associations, Strasbourg), and was its president in 1987-1988 and 1998-2000. In May 2000 she was unanimously elected president emeritus. She was a member of the National Council of Italian Journalists (1992-1998). From 2002 to 2004 she was member of the working group for scientific communication of the National Committee for Biotechnology. She has also been a consultant at the Italian Ministry of Research and Technology and editor-in-chief of the publication MRST, policy of science and technology. She has co-authored many publications in the field of scientific information, including Le biotecnologie in Italia, Le piste della ricerca and Luna vent’anni dopo.

Tags: tissue culture, domes, cells, epithelial cells

Duration: 3 minutes, 27 seconds

Date story recorded: May 2005

Date story went live: 24 January 2008