Culture of bovine embryos for stem cell production

THERIOGENOLOGY

CULTURE K.

OF

BOVINE

Schellander,

EMBRYOS

C.

and Department

of

The

aim

of

co1

out.

In

lected

Modified

ml

two

uere

were C

acids

amino purchased calf

recovered

the

conditions

were

the

experiment

first

compact to

6

2-4

mm

culture

FOUr

layer.

ICn

A

size

experiments,

was

embryos

in

attached

by

the

component

After

was

the

growth,

prevention

of Stage

a

ICM

culture

methods

of

10

the

the

cells.

The

cell

mass.

explantation, bovine

feeder

no stem

in

developed the

of experiment

efficient

to

two

incubation

fourth

stable

adhesion

or

next

non-essential

compact

further

small one

percent

and

Cell

the

first

cell

for layer

and

differentiation.

embryo

develovment

DHEH and

with

nucelosides

nonessent.

Exp.

1

DEEM without

amino Exp.

acids 2

and

nucleosides

nonessent. Exp.

3

amino

acids Exp.

4

0

compactmorula

4

6

a

5

24-72

hatching

3

6

7

3

72-144

growing

3

6

7

2

120-192

attachment

4

2

1

254

6

feeder of

the

and

min

were be

the

and

In

dark,

For

to

trophoblast

cell

for

of

cell

trophoblast

small,

first

from

of

without

of

the

of

initiation

Eighty-seven

used

need

in

the

3

(HB)

cytoplasm

in

of

Uithin

a

of

60%

In 100%

after

stock

observed.

conditions

enhanced

(h)

the

of

Culture

Culture

as

humidity.

surface

prepared

and

were incubation

culture.

observed.

39-c.

mitomycin

mechanically

the

spreading

visible

development

step.

to

high

representing

“as

non-

reagents

blastocysts

isolated

nucleoside

continuous was

development

urn

on

occurred

be

1

The

of

days

amount

experiment

trypsination

futher

time

medi

third

days

on

ml

experiment

hatched

were

serum,

fibroblasts

and

7

could

increased

the

3

minimal

ui thout

and

second

phenotype,

nucleoli

Al

animals. air

ml

stock

cultured

95%

culture

nucleus,

culture acids

temperature

ICH

cell-like

large prominent

amino

in

calf

1

Calf

attachment

(ICM)

fetal

stock.

experiment,

masses

80

antibiotic

the

into

no

ml

were

after

second

attached

stem

with

several

the

cell

inner

HB.

but

In

in

developed

diameter,

experiment.

10

old

ly

containing

ml

layers. week

were

nonsurgical

mercaptoethanol,

C02,

hatching

embryos

experiments

medium

embryos

and

isolating

of

day-7

nucleoside

12

5%

75%

were

all

of

culture

different goal Four

1

ml

The

with

test the

(DIIEII),

feeder

testes

37’C

morulae days

Sigma.

Korb

Austria

cells.

ml

1

fibroblart

from

to

Sar”m,

and

from

treated

was

in

1

H.

Genetics,

experiments,

Medium

calf

(penicillin/streptomycin), essential

Vienna,

cultured

Eagle

newborn

and

of

stem

first

PRODUCTION

Fiihrer,

leger

with

embryonic the

embryos

Dulbeccols 10

morulae

bovine

carried

CELL

Breeding

investigation bovine

STEM F.

Sch

University

this

for

pluripoteotial

U.

Animal

Veterinary

methods

FOR

Hassan-Hauser,

blastocyst

JANUARY

1989 VOL. 31 NO. 1