(i) obtaining inner cell mass (ICM) cells
from a bovine or porcine blastocyst or
lCM progenitor cells from a bovine or porcine
preblastocyst stage embryo;
(ii) culturing said lCM or lCM progenitor cells on a feeder layer
culture
to produce multilayer cell colonies;
(iii) identifying from among the cells contained in the cultured
lCM or lCM progenitor cell colony those cells which exhibit
the following properties:
(a) small cytoplasmic/nuclear volume ratio ranging
from about 10/90 to 50/50;
(b) cytoplasmic vesicles;
(c) small individual cells ranging from about 10 to 20
pro in diameter;
(iv) separating one or a cluster of said identified cells from the rest
of the cell colony; and
(v) passaging said separated lCM or lCM progenitor cells onto another
feeder layer culture and
maintaining physical contact between the feeder cell layer
and the separated cell or cell cluster throughout the
entire culturing period
to produce CICM cells that may be incorporated into a bovine
or porcine embryo to form a chimeric bovine or
porcine.
F 12. Claim 44 represents Infigen's claims to the invention:
44. A method for producing a cultured ungulate embryonic cell, the
method comprising:
(a) obtaining one or more blastomeres from a preblastocyst or blastocyst stage
ungulate embryo;
(b) culturing said one or more blastomeres on a feeder cell layer to
produce one or more cell colonies;
(c) identifying from amongst the cells contained in one or more cell
colonies those cells exhibiting the following characteristics:
(i) a cytoplasmic:nuclear volume ratio of 50:50 or less;
(ii) cytoplasmic vesicles; and
(iii) a diameter ranging from about 15[tm to about 21 [Lm;
(d) separating one or more cells having said characteristics from the rest
of the cell colonies to provide one or more separated cells;
and
(e) passaging said one or more separated cells
onto a feeder cell layer and
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