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Appendix B: Animal Reproductive Cloning Data Tables on Reproductive Cloning Efficiency and Defects
Pages 111-143

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From page 111...
... , 244 sheep embryos were created using somatic cell nuclear transplantation techniques. The donor nuclei were taken from epitheliallike cells grown from a culture of embryonic stem cells (Column 2~.
From page 112...
... , 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.
From page 113...
... Tables and Figures
From page 114...
... Embryonic 239 42 (18) N/A N/A stem cell Fetal fibroblast 276 33 (12)
From page 115...
... DATA TABLES ON CLONING EFFICIENCY AND DEFECTS 8 9 10 115 11 12 # Offspring # Live # Live births/ alive or healthy births/ # Embryos # Live births/ at time of Total# transferred to #Embryos publication/ Phenotypes fetuses(%) " uterus(%)
From page 116...
... >17 >16 Fetal fibroblast 174 35 (20) >3 >1 Adult skin cell 175 N/A 1 0 (0)
From page 117...
... DATA TABLES ON CLONING EFFICIENCY AND DEFECTS 8 9 10 117 11 12 # Offspring # Live # Live births/ alive or healthy births/ # Embryos # Live births/ at time of Total# transferred to #Embryos publication/ Phenotypes fetuses(%) " uterus(%)
From page 118...
... aThe species of animal used in the experiment. bThe cell type used as the source of the donor nucleus for the nuclear transfer.
From page 119...
... DATA TABLES ON CLONING EFFICIENCY AND DEFECTS 8 9 10 119 11 12 # Offspring # Live # Live births/ alive or healthy births/ # Embryos # Live births/ at time of Total# transferred to #Embryos publication/ Phenotypes fetuses(%) " uterus(%)
From page 120...
... Adult somatic cell nuclear transfer is used to preserve the last surviving cow of the Enderby Island cattle breed. Reprod.
From page 121...
... DATA TABLES ON CLONING EFFICIENCY AND DEFECTS # Fetuses after embryo transfer (pregnancies) #Live births # Offspring alive at time of publication 450 400 350 300 250 # 200 15010050 o Sheep Cattle Species 121 Mouse FIGURE 1 Survival Rates of Sheep, Cattle and Mouse Embryos Cloned from Adult, Fetal and Embryonic Cells.
From page 122...
... 122 SCIENTIFIC AND MEDICAL ASPECTS OF HUMAN REPRODUCTIVE CLONING 30 25 20 # 1510 5 o # Fetuses after embryo transfer (pregnancies) #Live births # Offspring alive at time of publication Goat Pig Monkey Species FIGURE 2 Survival Rates of Goat, Pig and Monkey Embryos Cloned from Adult, Fetal and Embryonic Cells.
From page 123...
... DATA TABLES ON CLONING EFFICIENCY AND DEFECTS 123 # Embryos created 8000 7000600050004000300020001 000 # Embryos developed into morula/ blastocyst #Embryos transferred # Fetuses after embryo transfer #Live births # Offspring alive at time of publication Sheep Cattle Mouse Goat Pig Monkey Species FIGURE 3 Efficiency of Cloning from Adult, Fetal and Embryonic Cells in Six Species.
From page 124...
... 124 SCIENTIFIC AND MEDICAL ASPECTS OF HUMAN REPRODUCTIVE CLONING TABLE 2 Phenotypes Observed in Cloned Animals 1 2 3 4 Speciesb Cell types # Live Defects seen in miscarried fetuses birthse Sheep Embryo-derived N/A 5 epithelial-like Adult mammary gland Fetal fibroblast Embryo-derived epithelial-like Fetal fibroblast ES cell line-derived epithelial-like injected into in vivo-matured oocytes N/A 1 2 fetuses from one of the cell types 3 showed abnormal liver development N/A 1 died after delayed delivery, 2 died after sibling (2) died in utero, 2 stillborn 3 late aborted fetuses underdeveloped for age; edema, hydronephrosis, testicular hypoplasia; also fetuses had variety of other defects, including cleft palate and interventricular septal defect 4 7 2 ES cell line-derived 1 late aborted fetus 1 epithelial-like underdeveloped for age; edema, injected into hydronephrosis, testicular in vitro-matured hypoplasia oocytes ES cell line-derived N/A 3 epithelial like ES cell line-derived N/A 3 fib rob last-like ES cell line-derived N/A 7 fib rob last-like Fetal fibroblast N/A 14
From page 125...
... 2/3 healthy and fertile; 1/3 died perinatally with respiratory failure 1/3 healthy; 1 died perinatally with respiratory failure, 1 died after being trampled by mother 2/7 healthy; 5/7 died with respiratory failure, 4 of those also had kidney problem (hydronephrosis) 3/14 alive, healthy; 7/14 died within 30 furs; 4/7 died within next 12 weeks: those that died had variety of unspecified kidney, liver and brain defects N/A N/A N/A N/A N/A N/A Campbell 1996 Wilmut 1997 Wilmut 1997 Wilmut 1997 Schnieke 1997 Wells 1997 necrosing placenta Wells 1997 N/A N/A N/A N/A Wells 1998h experiment 1 Wells 1998h experiment 2 Wells 1998h experiment 3 McCreath 2000 continues
From page 126...
... ; on necropsy, fetus was oversized and had abnormal lungs and heart Adult mural granulosa 1 case late miscarriage attributed 2 from a 13 yr old cow by researchers to hydrallantois 9 4 4 Adult cumulus N/A Adult oviduct N/A epithelial 5 3 Adult mural granulosa 7 miscarriages attributed by 10 researchers to hydrallantois
From page 127...
... , although these abnormalities did not compromise fetal development according to assessment of researchers continues
From page 128...
... 128 SCIENTIFIC AND MEDICAL ASPECTS OF HUMAN REPRODUCTIVE CLONING TABLE 2 Continued 1 2 3 4 Speciesb Cell types # Live Defects seen in miscarried fetuses birthse Adult mammary gland epithelium Adult ear skin fibroblast Fetal germ cell N/A Fetal fibroblast N/A Adult skin cell from N/A ES cell clone Adult muscle N/A 1 induced abortion at late gestation 1 due to hydrallantois: fetus oversized, cysts in kidney and liver, enlarged umbilical vessels 1 induced abortion at late gestation 1 due to hydrallantois: fetus oversized, cysts in kidney and liver, enlarged umbilical vessels 1 2 1 4 Fetal fibroblast 1/5 from miscarriage at 8 weeks; 8 4/5 from mothers that died late in pregnancy: 2/5 had chronic pulmonary hypertension and placental edema Senescent adult N/A fibroblast 6
From page 129...
... had astasia associated with arthrogryposis (abnormally developed joints) ; all cloned calves had high birth weight 5/8 were normal at birth, but 1/5 of these died at 6 weeks with suspected dilated cardiomyopathy; 3/8 had neonatal respiratory problems resulting in 1 death at 4 days; 2/8 were hydrallantoic pregnancies and only 1/2 of these survived; birth weights of all calves normal 6/6 had increased birth weight and some N/A born with moderate polyuria/ polydypsia, several had pulmonary hypertension and respiratory distress at birth, some had fever following vaccination N/A both calves that died had edematous placentas Zakhartchenko l999k Zakhartchenko l999k Zakhartchenko 1999L Zakhartchenko l999m Renard 1999 Shiga 1999 Hill 1999 Lanza 2000n continues
From page 130...
... Adult cumulus N/A 31 ES cells and mural N/A trophectoderm Adult fibroblast N/A 4 3
From page 131...
... , low CD45 antigen expression (required for T cell activation) N/A Kato 2000 2/6 placentas examined Hill 2000° from cloned pregnancies were normal; 4/6 were abnormal: 2/6 had flat cuboidal chronic epithelium and decreased vascularity; 2/6 had diminished cotyledonary structure N/A Hill 2000P phenotypes not described N/A Cheong 1993 6/25 (identical septuplet males)
From page 132...
... 132 SCIENTIFIC AND MEDICAL ASPECTS OF HUMAN REPRODUCTIVE CLONING TABLE 2 Continued 1 2 3 4 Speciesb Cell types # Live Defects seen in miscarried fetuses birthse Immature adult N/A Sertoli cells Tail tip fibroblast Adult cumulus Adult cumulus N/A Fetal fibroblast N/A Embryonic stem cell N/A all miscarriages were early in 7 pregnancy N/A 5 28 Goat Blastomere N/A 45 (embryonic) Fetal fibroblast all miscarriages were early in 3 pregnancy Fetal fibroblast N/A 6 infections 16 35 5 tested (does not say how many were born)
From page 133...
... compared to controls but researchers suggest this may have been caused by the agouti gene in their background 3/5 normal and healthy; 2/5 died with umbilical hernia and respiratory deficiency 28/28 had high birth weights, but this did not adversely affect clones in terms of survival; no respiratory or other problems 45/45 healthy N/A Wakayama 2000 Tamashiro 2000 placental hypertrophy Ono 2001 and also placental structural abnormalities high placental weights Eggan 2001 N/A Yong 1998 3/3 normal, healthy N/A Baguisi 1999 3/6 healthy; 3/6 died with respiratory placentas within normal Keefer 2001 infections range for # of cotyledons continues
From page 134...
... Wells et al. Adult somatic cell nuclear transfer is used to preserve the last surviving cow of the Enderby Island cattle breed.
From page 135...
... DATA TABLES ON CLONING EFFICIENCY AND DEFECTS 135 6 7 Placental defects, Phenotypes of five born clonesf phenotypes" Referencea 5/5 pigs very healthy N/A Polejaeva 1/1, Xena, is healthy no phenotypes described 2000 normal placenta Onishi 2000 N/A Betthauser 2000 2/2 healthy N/A Meng 1997
From page 136...
... Mouse Zygote Zygote Zygote 2-cell blastomere 2-cell blastomere Mll oocyte Inner cell mass Trophectoderm Pronuclear 2-cell 4-cell 8-cell Inner cell mass 8-cell Inner cell mass 8-cell 2-cell 8-cell Cumulus cell 4-cell 8-cell Inner cell mass 2-cell 8-cell Inner cell mass 2-cell 4-cell 8-cell 4-cells Inner cell masse TrophectodermC ES cell ES cell 16% (23/142)
From page 137...
... 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.
From page 138...
... Nat Genet 2000; 24:108-9. CAchieved using serial nuclear transfer.
From page 139...
... Zakhartchenko Fetal germ cells can direct development to term. 1999 Stice 1988 First production of genetically verified nuclear transfer rabbits from embryonic donor nuclei.
From page 140...
... Transgenic Fetal fibroblast Transgenic Fetal fibroblast Cattle Mll oocyte Cumulus cell Fetal fibroblast 0 (0/91)
From page 141...
... Serial nuclear transfer of cultured follicular cells but not cumulus cell nuclei results in term development. Male-derived adult somatic cell nuclei can direct term development; Immature, actively dividing Sertoli cell nuclei can direct term development.
From page 142...
... Production of male cloned mice from fresh, cultured and cryopreserved immature Sertoli cells.
From page 143...
... 0 Collas and Rob, First production of genetically verified unpublished nuclear transfer rabbits.


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