The prevalence of maternal F cells in a pregnant population and potential overestimation of foeto-maternal haemorrhage as a consequence.

BACKGROUND: Acid elution (AE) is used to estimate foeto-maternal haemorrhage (FMH). However AE cannot differentiate between cells containing foetal or adult haemoglobin F (F cells), potentially leading to false positive results or an overestimate of the amount of FMH. The prevalence of F cells in pregnant populations remains poorly characterised. The purpose of this study was to ascertain the incidence of HbF-containing red cells in our pregnant population using anti-HbF-fluorescein isothiocyanate flow cytometry (anti-HbF FC) and to assess whether its presence leads to a significant overestimate of FMH. MATERIAL AND METHODS: Eighty-eight pregnant patients were assessed for the presence of F cells and foetal red cells by AE and anti-HbF FC. The "FMH equivalent", estimated by AE and anti-HbF FC, was calculated. RESULTS: Thirty-six percent of the pregnant population had F-cell populations detectable by anti-HbF FC while AE detected F cells in 48% of the population. The mean estimated FMH equivalent determined by AE and anti-HbF FC was 0.59 mL (0-23.93 mL) and 0.41 (0 to 2.19 mL), respectively (p=0.012). In 3% of our population, AE overestimated the FMH by >3 mL due to the presence of an F-cell population of at least 16%. DISCUSSION: Thirty-six percent of a prospectively evaluated group of consecutive pregnant women were found to have F-cell populations. In some patients, these findings were clinically significant as AE overestimated the degree of FMH as a consequence.

Temporal expression of transcripts related to embryo quality in bovine embryos cultured from the two-cell to blastocyst stage in vitro or in vivo.

The post-fertilization embryo culture environment can have a dramatic effect on the pattern of gene expression in the embryo and it is widely acknowledged that bovine embryos derived from in vitro culture are of inferior quality to those derived in vivo. The objective of this study was to examine temporal variation in the mRNA abundance of several transcription and translation factors known to differ between blastocysts produced following culture in vitro and in vivo. Embryos were recovered from two in vitro culture systems SOF1 or SOF2 at five developmental stages: 2- to 4-cell, 8-cell, 16-cell, morula, and blastocyst. In vivo embryos were produced from superovulated and artificially inseminated heifers and recovered at approximately 40 hr or 3, 4, 5, and 7 days postinsemination. Blastocysts were also produced following in vitro maturation, in vitro fertilization and culture in the ewe oviduct. Analysis of relative transcript abundance for FOXO3A, EEF1G, HMG2, and REA was performed using quantitative real-time PCR. Irrespective of culture environment each transcript followed, approximately the same general pattern of expression where relative abundance decreased dramatically from the 2- to 4-cell stage to 8-cell stage and increased from the morula to blastocyst stage (P < 0.05). Transcripts for GNBL2 were not observed between the 2- and 16-cell stage of development. Relatively high expression at the 2- to 4-cell indicated that these transcripts are most likely of maternal origin produced in the oocyte during growth and final maturation. A culture-induced change in mRNA abundance of transcription and translation factors was evident in embryos that were produced not only between in vivo and in vitro culture environments but also between different in vitro culture systems.

Predicting embryo quality: mRNA expression and the preimplantation embryo.

To overcome the low implantation rate (10-20%) following IVF in humans, more than two embryos are commonly replaced, potentially leading to high order multiple pregnancies with associated significantly elevated risks. Selecting the most viable embryos and transferring fewer of them could reduce this risk. Prolonged culture of embryos in vitro to the blastocyst stage may expose the embryo to hazards not normally encountered in the female reproductive tract. Recent studies comparing bovine oocyte maturation, fertilization and embryo culture in vivo and in vitro have demonstrated that the origin of the oocyte is the main factor affecting blastocyst yield, while the post-fertilization culture environment is crucial in determining blastocyst quality, measured in terms of cryotolerance and relative transcript abundance, irrespective of the origin of the oocyte. Production of embryos in vitro, particularly when using an extended period of in-vitro culture may predispose the embryo to phenomena such as 'large offspring syndrome', which is probably linked to altered gene expression, particularly of imprinted genes. Post-fertilization culture environment clearly has a profound effect on the relative abundance of gene transcripts within the embryo. Culture under sub-optimal conditions for even one day can lead to perturbations in the pattern of expression.