Nuclear transfer in cattle using colostrum-derived mammary gland epithelial cells and ear-derived fibroblast cells.
To assess the developmental potential of nuclear transfer embryos in cattle using mammary gland epithelial (MGE) cells derived from the colostrum, we compared the effectiveness of cloning using those cells and fibroblast cells derived from the ear. The fusion rate of the enucleated oocytes with fibroblast cells (75 +/- 4%) was significantly higher than that with MGE cells (56 +/- 7%, P<0.05). There were no significant differences in the cleavage rate (85 +/- 3% vs. 91+/- 2%) or in the developmental rate to the blastocyst stage (35 +/- 6% vs. 35 +/- 5%) using MGE cells vs. fibroblast cells as donor nuclei (P>0.05). After transfer of blastocysts derived from nuclear transfer embryos produced using MGE cells and fibroblast cells, 13% (4/31) and 16% (6/37) of recipient heifers were pregnant on Day 42 as assessed by ultrasonography, respectively. Two of the 4 and 4 of the 6 recipients of embryos with MGE cell- and fibroblast cell-derived nuclei, respectively, aborted within 150 days of pregnancy. Four live female calves were obtained from MGE cells or fibroblast cells. However, one died from internal hemorrhage of the arteria umbilicalis. The other three calves were normal and healthy. There were no differences in the pregnancy rate or calving rate when using MGE cells vs. fibroblast cells. Microsatellite DNA analyses confirmed that the cloned calves were genetically identical to the donor cows and different from the recipient heifers. We conclude that colostrum-derived MGE cells have the developmental potential to term by nuclear transfer, and the efficiency of development of those cloned embryos was the same as that of embryos obtained using fibroblast cells as donor nuclei, although there was a significant difference in the fusion rate. This method using MGE cells derived from colostrum, which is obtained easily and safely from live adult cows, is more advantageous for cloning with somatic cells.
Kishi
M
M
Embryo Transplantation Laboratory, Snow Brand Milk Products Co Ltd Tomakomai, Hokkaido, Japan.
Itagaki
Y
Y
Takakura
R
R
Imamura
M
M
Sudo
T
T
Yoshinari
M
M
Tanimoto
M
M
Yasue
H
H
Kashima
N
N
D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry; D002417:Cattle / Q000235:genetics / Q000502:physiology*; D018929:Cell Culture Techniques; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D004396:Coloring Agents / Q000737:chemistry; D003126:Colostrum / Q000166:cytology / Q000502:physiology*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D004431:Ear, External / Q000166:cytology / Q000502:physiology; D004624:Embryo Transfer / Q000662:veterinary; D004847:Epithelial Cells / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000502:physiology; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D008321:Mammary Glands, Animal / Q000166:cytology / Q000502:physiology*; D018895:Microsatellite Repeats / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000662:veterinary; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy
D001562:Benzimidazoles; D004396:Coloring Agents; D007633:Keratins; D004247:DNA; C017807:bisbenzimide ethoxide trihydrochloride