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Below are the first 10 and last 10 pages of uncorrected machine-read text (when available) of this chapter, followed by the top 30 algorithmically extracted key phrases from the chapter as a whole.
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Scientific and Medical of Aspects: Human Reproductive Cloning
B
Animal Reproductive Cloning Data Tables on Reproductive Cloning Efficiency and Defects
The purpose of these tables is to provide an overview of the data from animal cloning experiments done to date (August 2001). Table 1 describes the success/failure rates of reproductive cloning in animals, and Table 2 provides details of the defects or lack of observable defects in reproductively cloned animals. These data were obtained through a comprehensive review of the publications cited in the “Reference” column of each table. Only experiments that yielded live-born cloned offspring were included in the table.
Tables 1 and 2 developed by the panel are supplemented by Tables 3 and 4 developed by Lewis et al., 2001. Note that Tables 3 and 4 use the term “cytoplast” for what the panel calls “enucleated egg.”
How to read Table 1:
Example: The first line from the table can be read as following:
In the experiments described in the paper published by Campbell in 1996 (Column 12), 244 sheep embryos were created using somatic cell nuclear transplantation techniques. The donor nuclei were taken from epithelial-like cells grown from a culture of embryonic stem cells (Column 2). Of these 244 embryos, only 34, or 14%, went on to develop into the morula or blastocyst embryos that are used in the embryo transfer procedure (Column 4). All 34 of those developing embryos were transferred into the wombs of female sheep (as we can tell from Column 8, which indicates
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number of embryos transferred). Of those 34 embryos, only 8 individual pregnancies resulted (Column 5). Of those 8 pregnancies, 3, or 38%, ended in miscarriage, and 5, or 63%, went on to produce live offspring (Columns 6 and 7, respectively). Of the five lambs that were born alive, only 2 (40%) survived until the time of publication. In all, 2% of the 244 embryos created resulted in live offspring (Column 9), and 12.5% of the 34 embryos transferred into recipient female sheep resulted in live offspring (Column 8).
How to read Table 2:
Any given line in Table 2 gives an overview of the clinical outcomes of each animal reproductive cloning experiment. For example, in line 1, in the sheep nuclear transplantation experiments published by Campbell in 1996 (Column 7), no information was given concerning the defects seen in miscarried fetuses (Column 3) or about the characteristics of placentas from these pregnancies (Column 6). However, Columns 4 and 5 indicate that 2/5 of the cloned lambs produced in this experiment were healthy and normal, whereas 3/5 died of unknown causes.
Note about Figures 1, 2, and 3
Figures 1, 2, and 3 were generated based on data presented in Table 1. Certain experiments whose results are displayed in Table 1 were omitted from the graphs due to incomplete data for all categories displayed in the graphs. Data from reproductive cloning experiments using embryonic, fetal and adult cells as nucleus donors were included in these graphs.
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Tables and Figures
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TABLE 1 Rates of Success/Failure of Somatic Cell Nuclear Transfer in Mammals
1
2
3
4
5
6
Speciesa
Cell typeb
# Embryos producedc
# Embryos developed into morula/ blastocyst (%)d
# Fetuses after embryo transfere
# Fetuses miscarried (%)f
Sheep
Embryo-derived epithelial-like
244
34 (14)
8
3 (38)
Adult mammary gland
277
29 (12)
1
0 (0)
Fetal fibroblast
172
47 (27)
5
2 (40)
Embryo-derived epithelial-like
385
126 (33)
15
11 (73)
Fetal fibroblast
507
69 (13.6)
14
7 (50)
ES cell line-derived epithelial-like
128
31 (24.2)
>9
>7 (~78)
ES cell line-derived epithelial-like
258
44 (17)
>11
>10 (~91)
ES cell line-derived epithelial-like
423
75 (18)
8
5 (63)
ES cell line-derived fibroblast-like
158
39 (31)
10
7 (70)
ES cell line-derived fibroblast-like
187
51 (27)
15
8 (53)
Fetal fibroblast
417
80 (19)
20
6 (30)
Cattle
Blastomere (embryonic)
641
152 (24)
>13
>4 (~31)
Blastomere (embryonic)
132
84 (64)
N/A
N/A
Embryonic stem cell
239
42 (18)
N/A
N/A
Fetal fibroblast
276
33 (12)
6
2 (33)
Adult mural granulosa from 13 yr old cow
621
259 (42)
28
26 (93)
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7
8
9
10
11
12
# Live births/ Total # fetuses (%)g
# Live births/ # Embryos transferred to uterus(%)h
# Live births/ # Embryos produced (%)i
# Offspring alive or healthy at time of publication/ # Live births (%)j
Phenotypes observedk
ReferenceL
5/8 (63)
5/34 (12.5)
5/244 (2.0)
2/5 (40)
#
Campbell 1996
1/1 (100)
1/29 (3.4)
1/277 (.36)
1/1 (100)
#
Wilmut 1997
3/5 (60)
3/40 (7.5)
3/172 (1.7)
2/3 (67)
E#
Wilmut 1997
4/15 (27)
4/87 (4.6)
4/385 (1.0)
4/4 (100)
#
Wilmut 1997
7/14 (50)
7/67 (10.4)
7/507 (1.3)
5/7 (71)
BC#
Schnieke 1997
2/>9 (<22)
2/31 (6.5)
2/128(1.6)
2/2 (100)
CE#
Wells 1997 in vivo-matured oocytes
1?>11 (<9)
1/44 (2.3)
1/258 (.39)
0/1 (0)
BEF
Wells 1997 in vitro-matured ooctyes
3/8 (38)
3/75 (4.0)
3/423 (.7)
2/3 (67)
B#
Wells 1998n experiment 1
3/10 (30)
3/39 (7.7)
3/158 (1.9)
1/3 (33)
B#
Wells 1998n experiment 2
7/15 (47)
7/44 (16)
7/187 (3.7)
2/7 (29)
BE#
Wells 1998n experiment 3
14/20 (70)
14/80 (17.5)
14/417 (3.4)
3/14 (21)
E#
McCreath 2000
N/A
9/59 (15)
9/641 (1.4)
N/A
N/A
Chesne 1993
N/A
19/78 (24)
19/132 (14)
N/A
N/A
Cheong 1993
N/A
4/34 (12)
4/239 (1.7)
N/A
N/A
Sims 1994
4/6 (67)
4/28 (14.3)
4/276 (1.4)
3/4 (75)
ABCF#
Cibelli 1998
2/28 (7.1)
2/74 (2.7)
2/621 (.32)
1/2 (50)
CD#
Wells 1998o
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1
2
3
4
5
6
Speciesa
Cell typeb
# Embryos producedc
# Embryos developed into morula/ blastocyst (%)d
# Fetuses after embryo transfere
# Fetuses miscarried (%)f
Adult cumulus
47
18 (38)
5
0 (0)
Adult oviduct epithelial
94
20 (21)
3
0 (0)
Adult mural granulosa
552
383 (69)
45
35 (78)
Adult mammary gland epithelium
140
36 (26)
>2
>1
Adult ear skin fibroblast
82
49 (60)
>5
>1
Fetal germ cell
279
85 (30)
>17
>16
Fetal fibroblast
174
35 (20)
>3
>1
Adult skin cell from ES cell clone
175
N/A
1
0 (0)
Adult muscle
346
73 (21)
8
4 (50)
Fetal fibroblast
876
>110? (>13)
>36
>28 (~78)
Adult senescent fibroblast
1896
87 (4.6)
>18
>11 (~61), 1 inducedm
Adult fibroblast from 17 yr old bull
338
103 (30)
12
6 (50)
Many adult and fetal types
1502
596 (40)
>50
>26 (~52)
Adult and fetal fibroblast
N/A
N/A
>54
>50 (~92)
Adult fibroblast from 21 yr old bull
190
53 (28)
6
1 inducedm
Mice
Adult cumulus
2468
1385 (56)
N/A
N/A
Embryonic stem cell
36
23 (64)
N/A
N/A
Mural trophectoderm
26
16 (62)
N/A
N/A
Adult fibroblast
463
377 (81)
N/A
N/A
Immature adult Sertoli cell
1846
436 (24)
235
219 (93)
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7
8
9
10
11
12
# Live births/ Total # fetuses (%)g
# Live births/ # Embryos transferred to uterus(%)h
# Live births/ # Embryos produced (%)i
# Offspring alive or healthy at time of publication/ # Live births (%)j
Phenotypes observedk
ReferenceL
5/5 (100)
5/6 (83)
5/47 (11)
2/5 (40)
#
Kato 1998
3/3 (100)
3/4 (75)
3/94 (3)
2/3 (67)
#
Kato 1998
10/45 (22)
10/100 (10)
10/552 (1.8)
10/10 (100)
ABC#
Wells 1999
1/>2 (<50)
1/4 (25)
1/140 (.7)
1/1 (1)
#
Zakhartchenko 1999p
1/>5 (<20)
1/16 (6.3)
1/82 (1.2)
0/1 (0)
AG
Zakhartchenko 1999p
1/>17 (<6)
1/32 (3.1)
1/279 (.36)
0/1 (0)
N/A
Zakhartchenko 1999q
2/>3 (<67)
2/7 (29)
2/174 (1.1)
1/2 (50)
AB#
Zakhartchenko 1999r
1/1 (100)
1/6 (16)
1/175 (.57)
0/1 (0)
CD
Renard 1999
4/8 (50)
4/26 (15)
4/346 (1.2)
1/4 (25)
ABG#
Shiga 1999
8/36 (22)
8/110 (7.2)
8/876 (.9)
6/8 (75)
BCF#
Hill 1999
6/>18 (<33)
6/79 (7.6)
6/1896 (.32)
6/6 (100)
ABD#
Lanza 2000s
6/12 (50)
6/54 (11)
6/338 (1.2)
4/6 (67)
AD#
Kubota 2000
24/>50 (<48)
24/172 (14)
24/1502 (1.6)
13/24 (54)
ADEG#
Kato 2000
4/>54 (<7.4)
4/243 (1.6)
4/?
1/4 (25)
BCDF#
Hill 2000t
1/6 (17)
1/26 (3.8)
1/190 (.52)
1/1 (100)
BD#
Hill 2000u
N/A
31/1385 (2.2)
31/2468 (1.3)
22/31 (71)
#
Wakayama 1998
N/A
2/18 (11)
2/36 (5.6)
N/A
N/A
Tsunoda 1998
N/A
2/25 (8)
2/26 (7.7)
N/A
N/A
Tsunoda 1998
N/A
3/274 (1.1)
3/463 (.6)
1/3 (33)
BF#
Wakayama 1999
16/235 (6.8)
16/436 (3.7)
16/1846 (.87)
15/16 (94)
GF#
Ogura 2000v
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1
2
3
4
5
6
Speciesa
Cell typeb
# Embryos producedc
# Embryos developed into morula/ blastocyst (%)d
# Fetuses after embryo transfere
# Fetuses miscarried (%)f
Tail tip fibroblast
753
260 (41)
126
119 (94)
Adult cumulus
3920
N/A
N/A
N/A
Fetal fibroblast
938
278 (30)
45
40 (89)
Adult cumulus (from hybrid strains)
4326
2583 (60)
N/A
N/A
Embryonic gonadal cell
179
114 (64)
N/A
N/A
Embryonic stem cell (from hybrid strain)
783
169 (22)
N/A
N/A
Goat
Blastomere (embryonic)
354
96
N/A
N/A
Fetal fibroblast
230
89 (39)
20
17 (85)
Fetal fibroblast
198
157 (79)
>6?
N/A
Pig
Adult granulosa
>401
401 (?)
9
4 (44)
Fetal fibroblast
210
188 (90)
N/A
N/A
Fetal body cell
143
N/A
N/A
N/A
Fetal genital ridge
340
N/A
N/A
N/A
Monkey
Blastomere (embryonic)
78
59 (76)
3
1/3 (33)
A = High birth weight
B = Pulmonary problems
C = Cardiovascular abnormalities
D = Immune system abnormalities/infection
E = Kidney and/or liver abnormalities
F = Placental abnormalities
G = Joint malformations or other gross deformities
# = Healthy offspring produced
NOTE: “N/A” indicates that no data were available in the cited publication.
NOTE: ES cell = embryonic stem cell.
NOTE: (~ ) indicates percentages extrapolated from data available, as shown in other columns.
aThe species of animal used in the experiment.
bThe cell type used as the source of the donor nucleus for the nuclear transfer.
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7
8
9
10
11
12
# Live births/ Total # fetuses (%)g
# Live births/ # Embryos transferred to uterus(%)h
# Live births/ # Embryos produced (%)i
# Offspring alive or healthy at time of publication/ # Live births (%)j
Phenotypes observedk
ReferenceL
7/126 (5.6)
7/280 (2.5)
7/753 (.93)
7/7 (100)
#
Ogura 2000w
N/A
35/? (>.9%?)
35/3920 (.89)
34/35? (97?)
#
Wakayama 2000
5/45 (11)
5/272 (1.8)
5/938 (.53)
3/5 (60)
BGF#
Ono 2001
N/A
80/2573 (3.1)
80/4326 (1.8)
N/A
#
Wakayama 2001
N/A
6/114 (5.2)
6/179 (3.4)
5/6 (83)
#
Wakayama 2001
N/A
28/? (>16.6?)
28/783 (.36)
22/28 (79)
ABF#
Eggan 2001
45/?
45/141 (32)
45/354 (13)
N/A
#
Yong 1998
3/20 (15)
3/85 (3.5)
3/230 (1.3)
3/3 (100)
#
Baguisi 1999
N/A
6/97 (6.1)
6/198 (3.0)
3/6 (50)
#D
Keefer 2001
5/9 (55)
5/401 (1.2)
5/>401 (<1.2)
5/5 (100)
#
Polejaeva 2000
N/A
1/110 (.9)
1/210 (.5)
1/1 (100)
#
Onishi 2000
N/A
2/143 (1.4)
2/143 (1.4)
N/A
N/A
Betthauser 2000
N/A
2/164 (1.2)
2/340 (.59)
N/A
N/A
Betthauser 2000
2/3 (67)
2/29 (6.9)
2/78 (2.6)
2/2 (100)
#
Meng 1997
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cThe number of embryos that were successfully formed after the nuclear transfer (cloning) procedure (in the literature usually referred to as # “fused” or # “reconstituted”).
In cases where this number was not available, the total number of oocytes injected with nuclei was used (including both successful and failed attempts to produce embryos).
In cases of double (serial) nuclear transfer, numbers of successfully reconstructed embryos from the second transfer were used.
dThe number and percentage of cloned embryos that continued to develop past the one-cell stage into multicellular embryos called morulae or blastocysts.
fThe number of fetuses that were spontaneously aborted at any time during the pregnancy.
gThe proportion of pregnancies that were carried to term (comparison of # live births to total # pregnancies).
IThe proportion of cloned embryos that went on to become live offspring (comparison of live births to the number of cloned embryos created).
jThe survival rate of live born clones after birth (comparison of live born offspring to the number still alive at the time of publication of the reference from which the data were obtained).
kThe letters indicate categories of characteristics observed in cloned animals (miscarried, live born or those that died after birth). Categories are provided above, in a key located below the table.
LThe peer reviewed scientific article in which data for any given experiment were published. Full references can be found in the bibliography.
mFetal miscarriage (abortion) was induced by researchers for medical or research reasons.
nWells et al. Cloning sheep from cultured embryonic cells. Reprod. Fertil. Dev. 1998; 10:615-626.
oWells et al. Adult somatic cell nuclear transfer is used to preserve the last surviving cow of the Enderby Island cattle breed. Reprod. Fertil. Dev. 1998; 10:369-378.
pZakhartchenko et al. Adult cloning in cattle: Potential of nuclei from a permanent cell line and from primary cultures. Mol. Reprod. Fertil. 1999; 54:264-272.
qZakhartchenko et al. Potential of fetal germ cells for nuclear transfer in cattle. Mol. Reprod. Dev. 1999; 52:421-426.
rZakhartchenko et al. Effects of serum starvation and re-cloning on the efficiency of nuclear transfer using bovine fetal fibroblasts. J. Reprod. Fertil. 1999; 115:325-331.
sLanza et al. Extension of cell life-span and telomere length in animals cloned from senescent somatic cells. Science 2000 Apr 28; 288:665-669.
tHill et al. Evidence for placental abnormality as the major cause of mortality in first-trimester somatic cell cloned bovine fetuses. Biol. Reprod. 2000; 63:1787-1794.
uHill et al. Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells. Biol. Reprod. 2000; 62:1135-1140.
vOgura et al. Production of male cloned mice from fresh, cultured, and cryopreserved immature Sertoli cells. Biol. Reprod. 2000; 62:1579-1584.
wOgura et al. Birth of mice after nuclear transfer by electrofusion using tail tip cells. Mol. Reprod. Dev. 2000; 57:55-59.
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FIGURE 1 Survival Rates of Sheep, Cattle and Mouse Embryos Cloned from Adult, Fetal and Embryonic Cells.
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5
6
7
Phenotypes of live born clonesf
Placental defects, phenotypesg
Referencea
15/16 pups normal; 1/16 had umbilical hernia, but was viable at birth
unusually large but structurally normal placentas
Ogura 2000q
7/7 pups normal, healthy
N/A
Ogura 2000r
telomeres lengthened rather than shortened in successive generations suggesting no inherited aging problem, all mice tested normal for behaviors (learning, memory, activity, agility, strength)
N/A
Wakayama 2000
5 of the healthy cloned mice were tested for behavioral defects - 3/10 measures of preweaning development were delayed but did appear and had no long-term effects; cloned mice were normal for learning, memory, activity and motor skills - these mice had high postnatal weight gain (not heavy at birth as in LOS) compared to controls but researchers suggest this may have been caused by the agouti gene in their background
N/A
Tamashiro 2000
3/5 normal and healthy; 2/5 died with umbilical hernia and respiratory deficiency
placental hypertrophy and also placental structural abnormalities
Ono 2001
28/28 had high birth weights, but this did not adversely affect clones in terms of survival; no respiratory or other problems
high placental weights
Eggan 2001
45/45 healthy
N/A
Yong 1998
3/3 normal, healthy
N/A
Baguisi 1999
3/6 healthy; 3/6 died with respiratory infections
placentas within normal range for # of cotyledons
Keefer 2001
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1
2
3
4
Speciesb
Cell typec
Defects seen in miscarried fetusesd
# Live birthse
Pig
Adult granulosa
N/A
5
Fetal fibroblast
N/A
1
Body cell and genital ridge cell
N/A
4
Monkey
Blastomere (embryonic)
N/A
2
NOTE: N/A indicates that no data were available in the cited publication
NOTE: ES cell = embryonic stem cell.
NOTE: LOS = large offspring syndrome.
aThe peer reviewed scientific article in which data for any given experiment were published. Full references can be found in the bibliography.
bThe species of animal used in the experiment.
cThe cell type used as the source of the donor nucleus for the nuclear transfer.
dDescription of abnormalities seen in aborted cloned fetuses; in some cases, these abnormalities may be the cause of miscarriage.
eThe number of live-born cloned animals.
fDescription of observations of physical, physiological or genetic characteristics of live born cloned animals at time of publication of cited refernces, unless stated otherwise.
gDescription of any characteristics, normal or abnormal, noted in the placentas of live born or miscarried cloned animals.
hWells et al. Cloning sheep from cultured embryonic cells. Reprod. Fertil. Dev. 1998; 10:615-626.
ICibelli et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 1998; 280:1256-8.
jWells et al. Adult somatic cell nuclear transfer is used to preserve the last surviving cow of the Enderby Island cattle breed. Reprod. Fertil. Dev. 1998; 10:369-378.
kZakhartchenko et al. Adult cloning in cattle: Potential of nuclei from a permanent cell line and from primary cultures. Mol. Reprod. Fertil. 1999; 54:264-272.
LZakhartchenko et al. Potential of fetal germ cells for nuclear transfer in cattle. Mol. Reprod. Dev. 1999; 52:421-426.
mZakhartchenko et al. Effects of serum starvation and re-cloning on the efficiency of nuclear transfer using bovine fetal fibroblasts. J. Reprod. Fertil. 1999; 115:325-331.
nLanza et al. Extension of cell life-span and telomere length in animals cloned from senescent somatic cells. Science 2000 Apr 28; 288:665-669.
oHill et al. Evidence for placental abnormality as the major cause of mortality in first-trimester somatic cell cloned bovine fetuses. Biol. Reprod. 2000; 63:1787-1794.
pHill et al. Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells. Biol. Reprod. 2000; 62:1135-1140.
qOgura et al. Production of male cloned mice from fresh, cultured, and cryopreserved immature Sertoli cells. Biol. Reprod. 2000; 62:1579-1584.
rOgura et al. Birth of mice after nuclear transfer by electrofusion using tail tip cells. Mol. Reprod. Dev. 2000; 57:55-59.
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5
6
7
Phenotypes of live born clonesf
Placental defects, phenotypesg
Referencea
5/5 pigs very healthy
N/A
Polejaeva 2000
1/1, Xena, is healthy
normal placenta
Onishi 2000
no phenotypes described
N/A
Betthauser 2000
2/2 healthy
N/A
Meng 1997
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TABLE 3 Developmental Capacity of Cytoplasts Reconstituted with Nuclei from Embryonic Cells
1
2
3
4
5
Species
Recipient cytoplast
Donor cell type
% Early Development: % Blastocyst (# Blastocysts/ # Cultured)
% Term Development: % Offspring (# Live births/ # Transferred)
Mouse
Zygote
Inner cell mass
16% (23/142)
19% (3/16)
Trophectoderm
1% (1/68)
0
Zygote
Pronuclear
95% (20/21)
no transfer
2-cell
13% (19/151)
no transfer
4-cell
0 (0/81)
no transfer
8-cell
0 (0/111)
no transfer
Inner cell mass
0 (0/84)
no transfer
Zygote
8-cell
0(0/32)
no transfer
Inner cell mass
0 (0/84)
no transfer
Zygote
8-cell
0(0/32)
no transfer
2-cell blastomere
2-cell
93% (40/43)
24%a (10/41)
Zygote
8-cell
51% (45/89)
0a (0/11)
Cumulus cell
0 (0/91)
no transfer
2-cell blastomere
4-cell
72% (49/68)
22% (10/46)
8-cell
35% (49/139)
8% (4/48)
Inner cell mass
0 (0/91)
no transfer
Mll oocyte
2-cell
23% (20/88)
15% (3/20)
8-cell
4% (1/26)
0 (0/1)
Inner cell mass
13% (11/87)
0 (0/11)
2-cell
78% (36/46)
29% (10/34)
4-cell
71% (30/42)
22% (6/27)
8-cell
46% (18/39)
18% (3/17)
4-cellc
83% (58/70)
43% (2/58)
Inner cell massc
64% (23/36)
11% (2/18)
Trophectodermc
62% (16/26)
8% (2/25)
ES cell
5% (47/931)
0 (0/56)
ES cell
29% (312/1087)
6% (8/132)
Sheep
Mll oocyte
8-cell
33% (8/24)
75% (3/4)
16-cell
27% (13/49)
21% (3/14)
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7
References
Significant findings
Illmensee 1981
First demonstration of developmental potential in mammals.
Reproducibility of results questioned.
McGrath 1984
Biologically impossible to achieve development with transcriptionally active nucleus.
Robl 1986
Development more advanced with cytoplast prepared from 2-cell than zygote.
Robl 1986
Development more advanced with cytoplast prepared from 2-cell than zygote.
No development beyond 12 days gestation.
Wakayama 2000b
No development when zygotic cytoplasts were used.
Robl 1987
Term development when 4- and 8-cell nuclei used but not more advanced.
Importance of cytoplast environment.
Kono 1991g
Development to term from embryonic nuclei transferred to enucleated oocyte.
Cheong 1993
Embryonic nuclei in G1 phase of the cell cycle can direct term development when transferred to Mll cytoplasts.
Kwon 1996
Serial nuclear transfer of metaphase-arrested embryonic nuclei results in term development.
Tsunoda 1998
Term development following serial nuclear transfer of inner cell mass and trophectoderm nuclei.
Tsunoda 1993
Implantation sites but no term development.
Wakayama 1999f
Late-passage actively dividing ES cell nuclei are able to direct development to term.
Willadsen 1986
Term development from cleavage stage blastocysts.
Smith 1989
Transcriptionally active nuclei are able to direct development to term.
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1
2
3
4
5
Species
Recipient cytoplast
Donor cell type
% Early Development: % Blastocyst (# Blastocysts/ # Cultured)
% Term Development: % Offspring (# Live births/ # Transferred)
Inner cell mass
38% (6/16)
11% (1/9)d
Cultured cell line
14% (34/244)
14% (5/34)
Cattle
Pronuclear
Pronuclear
13% (5/38)
100% (2/2)
2- to 8-cell
0 (0/10)
no transfer
Mll oocyte
2- to 8-cell
12% (13/111)
0 (0/12)
9- to 16-cell
16% (8/50)
28 (2/7)
17- to 32-cell
8% (2/24)
no transfer
Morula (64-cell)
23-35%
22%e (104/463)
Morula (31-cell)
24% (152/641)
15% (9/59)
Inner cell mass
7% (20/304)
13% (2/15)
Inner cell mass
5% (30/629)
8% (2/26)
Cultured inner cell mass
27% (109/406)
12% (4/34)
Fetal germ cell (PGC)
20-38% (30/149-53/140)
5% (1/20)
Rabbit
Mll oocyte
8-cell
not assessed
4% (6/164)
8- to 16-cell
49% (34/69)
21% (23/110)
32-cell
33% (14/43)
15% (10/67)
Inner cell mass
20% (17/83)
no transfer
Trophectoderm
0 (0/52)
no transfer
Pig
Mll oocyte
2-cell
9% (1/11)
0 (0/33)
4-cell
8% (7/83)
3% (1/34)
8-cell
19% (11/57)
0 (0/21)
Goat
Mll oocyte
Morulac
31% (18/57)
31% (45/141)
Monkey
Mll oocyte
8-cell
52%e (53/101)
4% (2/53)
aDevelopment assessed at 8.5 days post coitum.
bWakayama et al. Nuclear transfer into mouse zygotes. Nat Genet 2000; 24:108-9.
cAchieved using serial nuclear transfer.
dDevelopment assessed at 42 days of pregnancy.
eEmbryonic development assessed at the 2-cell stage prior to transfer.
fWakayama et al. Mice cloned from embryonic stem cells. Proceedings of the National Academy of Sciences, USA 1999; 96:14984-89.
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References
Significant findings
Campbell 1996
Nuclei from cell lines from embryonic discs are able to support development to term.
Robl 1987
Cleavage stage embryonic nuclei are unable to direct embryonic or term development when transferred to enucleated zygotes.
Prather 1987
Term development from transcriptionally active donor embryonic nuclei.
Bondioli 1990
Nuclei from morula stage embryos can direct midgestation development.
Chesne 1993
Nuclei from morula stage embryos can direct development to term.
Collas 1994
Direct injection of inner cell mass nuclei into Mll cytoplasts can direct development to term.
Keefer 1994
Totipotency of inner cell mass nuclei confirmed.
Sims 1984
Nuclei from inner cell mass cultured for up to 28 days are able to direct development to term.
Zakhartchenko 1999
Fetal germ cells can direct development to term.
Stice 1988
First production of genetically verified nuclear transfer rabbits from embryonic donor nuclei.
Prather 1989
High rates of development from transcriptionally active embryonic nuclei.
Collas 1990; Callas 1991
Normal embryonic development from inner cell mass donor nuclei.
Prather 1989
Cleavage stage embryonic nuclei can direct term development in pigs.
Yong 1998
Serial nuclear transfer of transcriptionally nuclei results in high rates of development.
Meng 1997
Embryonic nuclei can support term development in the monkey.
gKono T et al. Development of enucleated mouse oocytes reconstituted with embryonic nuclei. J. Reprod. Fertil. 1991; 93:165-72.
SOURCE: Lewis, IM, MJ Munsie, AJ French, R Daniels and AO Trounson, 2001. The Cloning Cycle: From Amphibia to Mammals and Back. Reproductive Medicine Reviews 9:1 pp. 3-33.
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TABLE 4 Developmental Capacity of Cytoplasts Reconstituted by Nuclei from Fetal and Adult Somatic Cells
1
2
3
4
Species
Recipient cytoplast
Donor Cell Type
% Early Development: % Blastocyst (# Blastocysts/ # Cultured)
Mouse
Zygote Mll oocyte
Cumulus Cell Thymocyte
0 (0/91) 7% (6/88)
Cumulus Cell
67% (101/151)
Neuronal Cell
22% (50/223)
Sertoli cell (mature)
40% (63/159)
Cumulus Cell
20% (19/93)
Cultured follicular cell;
34% (51/151) 3% (1/30)
Adult Male fibroblast;
50% (207/414)
Cumulus cell.
52% (206/393)
Fibroblast cell
23% (38/162)
Sertoli Cell (immature)
33% (94/284)
Sheep
Mll oocyte
Fetal fibroblast
27% (34/124)
Adult Mammary (epithelial)
12% (29/247)
Transgenic Fetal fibroblast
5-21% (5/82-19/89)
Transgenic Fetal fibroblast
6-28% (14/109, 43/154, 4/71, 19/83)
Cattle
Mll oocyte
Cumulus cell
13% (5/38)
Fetal fibroblast
12% (33/276)
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7
% Term Development: % Offspring / (# Live births/ # Transferred)
References
Significant Findings
No Transfer 0
1. Wakayama 2000a -same footnote information as earlier Wakayama 2000 in Table 3
2. Callas, 1992
1. No development when zygotic cytoplasts were used.
2. Somatic nuclei are able to direct embryonic development through no term development.
2% (31/1315)
Wakayama 1998b
Direct-injected cumulus cell nuclei direct term development; however, Sertoli and neuronal nuclei do not.
2%a (1/46)
Findings do not support the requirement of G0/G1 nuclei for term development.
2%a (1/59) 0 (0/3)
Kato 1999
Serial nuclear transfer of cultured follicular cells but not cumulus cell nuclei results in term development. nuclei can direct term development.
3% (1/30);
Wakayama 1999c;
Male-derived adult somatic cell nuclei can direct term development; Immature, actively dividing Sertoli cell
1% (2/177);
Ogura 2000d
1%c (2/206)
0c (0/38)
4% (6/134)
8% (3/40)
Wilmut 1997
Inducing cell to enter quiescence by serum starvation may assist in nuclear reprogramming.
3% (1/29)
First demonstration that nuclei from differentiated somatic fetal or adult origin can direct development to term.
5/21% (1/21-1/5)
Schnieke 1997
Term development of transfected somatic cell nuclei suggests an alternative method for the production of transgenic animals. One male lamb was born.
0-28% (4/14, 8/43, 0/4, 2/19)
McCreath 2000
Production of gene-targeted sheep by nuclear transfer from cultured somatic cells.
0 (0-19)
Callas 1994
Nuclei from adult somatic cells can direct embryonic development.
14% (3/28)
Cibelli 1998e
Cultured activity dividing fetal fibroblast nuclei can direct development to term.
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2
3
4
Species
Recipient cytoplast
Donor Cell Type
% Early Development: % Blastocyst (# Blastocysts/ # Cultured)
Adult Male Fibroblast
21-37% (24/114 - 43/115)
Cumulus Cell
49% (18/37)
Cultured Oviductal cell
23-34% (196/842 - 29/84)
Cultured Granulosa
28% (152/552)
Rabbit
Mll oocyte
Adult granulosa
Number not specified
Pig
Mll oocyte
Granulosa cell line
Not assessed
Fetal fibroblast
1-31% (total 88/615) 93% (2-, 4- and 8-cell, 110/118)
Goat
Mll oocyte
Transgenic Fetal fibroblast
Not assessed
Monkey
Mll oocyte
Fetal fibroblast
57% (57/100)
Adult fibroblast
44% (4/9)
SOURCE: Lewis, IM, MJ Munsie, AJ French, R Daniels and AO Trounson, 2001. The Cloning Cycle: From Amphibia to Mammals and Back. Reproductive Medicine Reviews 9:1 pp. 3-33.
NOTE: Cytoplast = Enucleated Egg.
aWakayama et al. Nuclear Transfer into mouse zygotes. Nat Genet 2000; 24:108-9.
bWakayama et al. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 1998; 394:369-74.
cWakayama T. and Yanagimachi R. Cloning of male mice from adult tail-tip cells. Nat Genet 1999; 22:127-8
dOgura A, et al. Production of male cloned mice from fresh, cultured and cryopreserved immature Sertoli cells. Biol. Reprod. 2000; 62:1579-84.
eCibelli J., et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 1998; 280:1256-8.
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% Term Development: % Offspring / (# Live births/ # Transferred)
References
Significant Findings
7% (2/7)
Kubota 2000
Nuclei from male adult fibroblast can direct development to term.
33% (2/6)
Kato 1995
High rates of term development following transfer of cumulus and oviduct nuclei.
50% (2/4)
10% (10/100)
Wells 1999
Production of calves from cultured granulosa cells.
0
Collas and Rob, unpublished
First production of genetically verified nuclear transfer rabbits.
1.3% (5/401)
Polejaeva 2000
Term development following serial nuclei transfer of cumulus cells.
No Transfer 0.9% (1/110)
Onishi 2000
Term development following direct injection of nuclei from fetal fibroblast cells.
3% (3/112)
Baguisi 1999
Production of transgenic goats from transfected fetal fibroblast nuclei.
0
Wolf 1999
Donor nuclei from cell lines are capable of limited embryonic development.
0
Representative terms from entire chapter:
nuclear transfer