ABSTRACT
A utilização do soro fetal bovino (SFB), embora bastante disseminada na produção in vitro (PIV) de embriões bovinos, apresenta limitações por ser um meio indefinido e por causar efeitos que prejudicam a qualidade desses embriões. Por esse motivo, nos últimos anos, grande parte das pesquisas relacionadas à PIV está voltada para a substituição do SFB por outros compostos nos meios de cultura. No presente estudo, foram utilizados como compostos protéicos a albumina sérica bovina livre de ácidos graxos (BSA-FAF) e um produto comercial denominado fluido embriônico (FE) de maneira isolada ou em diferentes combinações e concentrações, com objetivo de substituir ou diminuir a concentração do SFB durante a maturação in vitro (MIV). [...] Ademais, o G3 também apresentou diminuição na taxa de maturação nuclear quando comparado ao G4. Quanto à maturação citoplasmática, nos grupos G2, G7, G6 e G3, houve redução (p<0,05) das taxas para 43,9por cento, 43,2 por cento, 43,1 por cento e 36,5 por cento, respectivamente, quando comparadas ao meio controle (G1), que permitiu a obtenção de valores médios de 62,4 por cento. Por outro lado, nos grupos G8, G4 e G5, a taxa de maturação citoplasmática não foi afetada com a redução do SFB, onde 59,3 por cento, 51,3 por cento e 50,8 por cento dos oócitos apresentaram os GC dispostos na periferia, respectivamente. Os resultados obtidos pelo teste de contrastes ortogonais complementam os obtidos na avaliação da maturação nuclear e migração de grânulos corticais, mostrando a necessidade do SFB durante a MIV, mesmo que em baixas concentrações, e a possibilidade de diminuir a sua concentração associando-o a BSA-FAF e/ou FE. Dessa forma, conclui-se que é possível reduzir a concentração de SFB no meio de MIV para até 3,5% sem prejuízo significativo aos índices de maturação nuclear e citoplasmática.
The use of fetal calf serum (FCS), although widely employed during in vitro production (IVP) of bovine embryos, has limitations. FCS is an undefined media and may have harmful effects on the quality of embryos. For this reason, in recent years, research efforts aimed at improving IVP of bovine embryos, have focused at the replacement of FCS by alternative compounds in culture media. In this study, fatty acid free bovine serum albumin (BSA-FAF) and embryonic fluid (EF) were used separately or in combination, in different concentrations, to replace or reduce the concentration of FCS during in vitro maturation (IVM). [...] Moreover, G3 also showed inferior nuclear maturation rate when compared to G4. Regarding cytoplasmic maturation, the rates were reduced to 43.9 percent, 43.2 percent, 43.1 percent and 36.5 percent in G2, G7, G6 and G3 groups, respectively, compared to the control group (G1; 62.4 percent). On the other hand, in the groups G8, G4 and G5, maturation rates were not affected by reduction of FCS, where 59.3 percent, 51.3 percent and 50.8 percent of the oocytes displayed CG arranged peripherally, respectively. The results obtained by the orthogonal contrast test are in accordance with the ones from the evaluation of the nuclear maturation and cortical granules migration. These data show the need of FCS on the MIV, even in low concentrations, and the possibility of decrease its concentration by associating it with BSA-FAF and/or EF. Therefore, we concluded that it is possible to reduce the concentration of FCS in IVM medium to a concentration of 3.5 percent without affecting nuclear and cytoplasmic maturation rates.
Subject(s)
Animals , Serum Albumin/genetics , Cattle/embryology , Serum Albumin, Bovine/genetics , In Vitro Oocyte Maturation Techniques/veterinary , Fertilization in Vitro/veterinary , Culture Techniques/veterinaryABSTRACT
Objective To investigate the effect of umbilical cord serum(UCS)during induction of dendritic cells derived from umbilical cord blood. Methods Umbilical cord blood was collected aseptically and interface cells were collected with density gradient centrifugation method,CD34 +cells were purified and collected with Mini-MACS technique. Pro-DC were obtained and were divided into 3 groups:①cultured in RPMI-1640 medium with 10%UCS and cytokine,②cultured in RPMI-1640 medium with 10%FCS and cytokine③cultured in RPMI-1640 medium with 10%FCS .Interleukin-4(IL-4),granulocyte-macrophage colony-stimulating factor(GM-CSF)and tumor necrosis factor(TNF-a)were used in the first 2 groups, but not in control group (group 3). Some cells were collected on the 8th d for phenotypes analysis. Results The expression of CD80 ,CD54 and HLA-DR on the surface of DC cultured in UCS medium had no significant difference compared with that of DC cultured in FCS medium. The expression of surface molecules in UCS-DC group and FCS-DC group were both enhanced compared with the control group. Conclusion Mature DC could be induced from pro-DC derived from umbilical cord blood when cultured with UCS instead of FCS. That provides a new approach for the clinical application of DC.
ABSTRACT
The abnormal proliferation of vascular smooth muscle cells after endothe-lial injury is postulated to be the main pathophysiological process in atherosclerosis (AS). The effects of propylene glycol mannate sulfate(PGMS) on the proliferation of bovine cerebral microvessel smooth muscle cslls (BCMSMCs) induced by 10% fetal calf serum (FCS) and interleukin l(IL-1) were investigated in culture. 5 - 8 stage subcultured BCMSMCs were incubated into 96-well dish- With either 10% FCS or IL-1 (50u/ml) to produce BCMSMCs proliferation , the inhibitory effects of PGMS on proliferation of BCMSMCs were investigated. The results shows that PGMS could inhibit the proliferation of quiescent BCMSMCs induced by 10% PCS. The growth of cells was inhibited, comparing with normal control 72hours after the serum addition as determined by crystal violet stainning and MTTmethod. The proliferation of quiescent BCMSMCs induced by IL-1 (50u/ml) was also inhibited by PGMS as determined by crystal violet stainning and MTT method. The results suggested that PGMS inhibit the proliferation of BCMSMCs induced by 10% FCS and IL-1 ,and the use of PGMS may probably play an important role in the treatment of cerebrovascular disease.
ABSTRACT
Conditions have been standardized to maintain rat vaginal epithelial cells in vitro with more than 95% viability. Cultured epithelial cells were used to study the effects of normal fetal calf scrum, estradiol and progesterone on the incorporation of [3H]-uridine in RNA and incorporation of [l4C]-aminoacids in proteins. While fetal calf serum and estradiol stimulate the incorporation of both uridine and afno acids, progesterone did not show any effect. Estradiol treated vaginal cells show typical fcroridges (indicative of keratinization of cells) in contrast to estradiol deprived cells, which show microvilli on cell surface when examined in scanning electron microscope.