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US 2003/0115622 A1Published application
Production of avian embryonic germ (eg) cell lines by prolonged culturing of pgc's, use thereof for cloning and chimerization
OwnerF. Abel Ponce De Leon, James M. Robl +2inventors
InventorsF. ABEL PONCE DE LEON, JAMES M. ROBL, STEVEN L. STICE, D. JOSEPH JERRY
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Abstract
A culture system for producing PGCs or EG cells by culturing PGCs for long periods in tissue culture is provided. This culture system uses LIF, bFGF, IGF and SCF. The resultant EG cells are useful for the production of transgenic and chimeric avians, in particular, chickens and turkeys, and also for cloning purposes.
Claims24
A culturing method which provides for the production of avian PGC and germ (EG) cells comprising the following steps:
- (i) isolating primordial germ cells from a desired avian;
- and
- (ii) culturing said primordial germ cells in a culture medium containing at least the following growth factors contained in amounts sufficient to maintain said PGCs for prolonged periods in tissue culture:
- (1) leukemia inhibitory factor (LIF),
- (2) basic fibroblast growth factor (bFGF),
- (3) stem cell factor (SCF) and
- (4) insulin-like growth factor (IGF), for prolonged time period sufficient to produce a culture having a compact multilayer like appearance;
- (iii) identifying EG cells contained therein.
5 more elements
An improved method of producing chimeric avians which comprises:
- (i) isolating primordial germ cells from an avian;
- (ii) maintaining such PGCs in a tissue culture medium containing at least the following growth factors;
- (1) leukemia inhibitory factor (LIF),
- (2) basic fibroblast growth factor (bFGF),
- (3) stem cell factor (SCF) and
- (4) insulin-like growth factor (IGF) for a sufficient time to produce embryonic germ (EG) cells;
- (iii) transferring said EG cells into a recipient avian embryo;
- and
- (iv) selecting for chimeric avians which have the desired PGC phenotype.
6 more elements
Dependencies are inferred from the claim text.