Effects of human versus mouse leukemia inhibitory factor on the in vitro development of bovine embryos.
Leukemia inhibitory factor (LIF) is a cytokine that shows conflicting effects on in vitro produced (IVP) bovine embryos. Bovine LIF (bLIF) has been cloned and used in culture, but there is no commercially available bLIF. Thus, researchers use human LIF (hLIF) to supplement the culture medium for bovine embryos because of its greater sequence homology compared to murine LIF (mLIF). We compared the effects of mLIF and hLIF on the development of bovine embryos in culture with the effects described for bLIF. Oocytes were matured and fertilized in vitro and cultured in modified synthetic oviduct fluid with BSA. On Day 6 post-insemination, morulae were cultured for 48h in the presence of: (1) mLIF, 100ngml(-1); (2) hLIF, 100ngml(-1); or (3) no LIF. Reduced blastocyst rates were observed on Day 8 for hLIF at the middle and expanded stages, while mLIF had no effect. In contrast, Day 8 blastocysts showed decreased cell counts both in terms of inner cell mass (ICM) and ICM/total cell proportions in the presence of mLIF, while hLIF had no effect. No changes were seen in trophectoderm (TE) and total cell counts. The increased hatching rates and TE cell counts previously described for bLIF, together with the disparate effects exhibited by hLIF and mLIF during blastocyst formation indicate these compounds are inappropriate to replace bLIF. We recommend that heterospecific LIF should not be used to supplement the culture medium for bovine embryo or embryonic stem cells.
Rodríguez
A
A
Géneticay Reproducción SERIDA, Camino de los Claveles 604, 33203 Gijón, Asturias, Spain.
De Frutos
C
C
Díez
C
C
Caamaño
J N
JN
Facal
N
N
Duque
P
P
García-Ochoa
C
C
Gómez
E
E
D000818:Animals; D002417:Cattle / Q000196:embryology*; D002452:Cell Count / Q000662:veterinary; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D006801:Humans; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D016014:Linear Models; D008297:Male; D051379:Mice
C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse