Processes for clonal growth of hepatic progenitor cells

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 5/071 (2010.01) C12Q 1/02 (2006.01) C12Q 1/18 (2006.01)

Patent

CA 2424779

A method for propagating mammalian endodermally derived progenitors such as hepatic progenitors, their progeny, or mixtures thereof is developed which includes culturing mammalian progenitors, their progeny, or mixtrues thereof on a layer of embryonic mammalian feeder cells in a culture medium. The culture medium can be supplemented with one or more hormones and other growth agents. These hormones and other growth agents can include insulin, dexamethasone, transferrin, nicotinamide, serum albumin, .beta.- mercaptoethanol, free fatty acid, glutamine, CuSO4 and H2SeO3. The culture medium can also include antibodies. Importantly, the culture medium does not include serum. The invention includes means of inducing the differentiation of the progenitors to their adult fates such as the differentiation of hepatic progenitor cells to hepatocytes or biliary cells by adding, or excluding epidermal growth factor, respectively. The method of producing mammalian progenitors is useful in that the progenitors can be used subsequently in one or more of the following processes: identification of growth and differentiation factors, toxicological studies, drug development, antimicrobial studies, or the preparation of an extracorporeal organ such as a bioartificial liver.

L'invention concerne un procédé permettant de propager des progéniteurs mammaliens d'origine endodermique, tels que les progéniteurs hépatiques, leur descendance, ou des mélanges de ceux-ci sur une couche de cellules nourricières embryonnaires mammaliennes dans un milieu de culture. Le milieu de culture peut être complété par une ou plusieurs hormones et par d'autres agents de croissance. Ces hormones et autres agents de croissance peuvent comprendre l'insuline, la dexaméthasone, la transferrine, la nicotinamide, la sérumalbumine, le .beta.-mercaptoéthanol, l'acide gras libre, la glutamine, le CuSO¿4? et le H¿2?SeO¿3?. le milieu de culture peut également comprendre des anticorps. Le milieu de culture ne comprend pas de sérum. Le procédé décrit dans cette invention comprend des moyens permettant d'inciter la différentiation des progéniteurs de leurs formes adultes, telle que la différentiation des cellules souches hépatiques des hépatocytes ou des cellules biliaires, par ajout ou par exclusion du facteur de croissance épidermique, respectivement. Le procédé de production de progéniteurs mammaliens est utile en ce que les progéniteurs peuvent être utilisés ultérieurement dans un ou plusieurs des procédés suivants: l'identification des facteurs de croissance et de différentiation, les études toxicologiques, la mise au point de médicaments, les études antimicrobiennes, ou la préparation d'un organe extra-corporel, tel qu'un foie bioartificiel.

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