Fetal losses and pathologic findings of clones derived from serum-starved versus serum-fed bovine ovarian granulosa cells

Abstract

The objective was to evaluate development of cloned embryos constructed with serum-starved versus fed ovarian granulosa cells. Fusion of somatic cells with cytoplasts (50.7 and 50.7%; SEM=5.5), development of cloned embryos to 8-16-cell (59.4 and 62.6%; SEM=6.4), and compact morula/blastocyst-stages (24.9 and 26.5%; SEM=8.5) was similar for serum-starved and fed groups. More heifers were confirmed pregnant with clones derived from serum-starved cells (9/13, 67.2% versus 11/25, 44%). However, embryonic loss between 29-50 days was greater for clones constructed with serum-starved (88.9%) versus fed (36.4%) cells. Development of clones derived from serum-fed cells through placentation and differentiation was not predictive of competency to term. Fetal deaths occurred in the majority of late term pregnancies as a result of complications from hydrallantois. Only one fetus derived from serum-fed ovarian granulosa cells developed to term (278 days). At birth and for approximately 9 consecutive months thereafter, routine veterinary examinations confirmed expected growth patterns. In summary, the use of serum fed granulosa cells for nuclear transfer was coincident with a high incidence of third trimester losses associated with hydrallantois and fetal oversize.