Ungulate embryonic stem-like cells, methods of making and...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/06 (2006.01) A01K 67/027 (2006.01) A61D 19/04 (2006.01) C12N 15/87 (2006.01)

Patent

CA 2242666

Primordial germ cells are extracted from post blastocyst embryos of an ungulate such as extracting primordial germ cells from the gonadal ridges of 25-day porcine embryos or 34-40 day bovine embryos. The primordial germ cells are cultured in long term culture (over 30 days) resulting in cells which resemble embryonic stem cells in morphology and with respect to maintaining pluripotency. The cells obtained can be maintained for several months in culture and can be genetically manipulated using homologous recombination technology in order to insert desired genetic material into the genetic compliment of the cell at a desired location. The genetically manipulated cell can be inserted into a blastocyst obtained from the same species of animal as the cell was derived in order to produce a chimeric ungulate which ungulate may, via its genetic engineering, produce desired pharmaceutical products.

On extrait des cellules germinales primordiales d'embryons post-blastocytes d'un ongulé, en prélevant, notamment, des cellules germinales primordiales dans les replis gonadiques d'embryons de porc âgés de 25 jours ou d'embryons de bovins dont l'âge se situe entre 34 et 40 jours. On cultive ces cellules germinales embryonnaires sur une longue période, plus de 30 jours, ce qui donne des cellules ressemblant, de par leur morphologie et le maintien de leur multipotence, à des cellules embryonnaires souches. Il est possible de maintenir en culture les cellules ainsi obtenues plusieurs mois durant et de les modifier par génie génétique en faisant intervenir des techniques de recombinaison homologue et ce, afin d'introduire un matériau génétique souhaité dans le complément génique de la cellule en un site désiré. On est en mesure d'introduire la cellule ainsi modifiée par génie génétique dans un blastocyte obtenu à partir d'un animal de la même espèce que celle chez qui a été dérivée la cellule afin de fabriquer un ongulé chimère, susceptible, du fait de sa mise au point par génie génétique, de produire des substances à usage pharmaceutique désirées.

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