[Are patients in the postpartum period potential egg donors?]. / Les accouchées sont-elles de potentielles donneuses d'ovocytes ?

OBJECTIVES: In France, oocyte donation program is still underdeveloped because of lack of donors and this situation entails an important wave of cross border medical tourism to different European countries mainly Spain and Greece. In 2011, the General inspection of social affairs report recommended to the biomedicine agency to promote spontaneous oocyte donation via different channels of information to develop this national program. The main objective of this study is to assess the knowledge of women after baby delivery about oocyte donation. The second objective is the identification of ways to assure better information and to promote oocyte donation. PATIENTS AND METHODS: We conducted a prospective study with anonymous questionnaire distribution to women after delivery at obstetrics/gynecology department of the Regional University Hospital and Maternity-Children Unit "Victor-Pauchet" of Amiens, from December 2012 to January 2013. RESULTS: Two hundred and fifty-five questionnaires were distributed and 242 of them were analyzed (94.9%). About oocyte donation knowledge: 28% did not know it was possible, 45% did not know it was legal in France, 54% did not know who was concerned and 36% know that a treatment is necessary, 9% think that oocyte donation is paid and 10% it is non-anonymous. If 67% seems to be favorable to this initiative, only 35% could accept to realize it. About information efficiency, 88% think not to receive enough information, 64% would like to have more information. The health care professional wanted to give this information is an obstetrician (51%), a midwife (37%) and a nurse (12%). DISCUSSION AND CONCLUSION: Oocyte donation program is misoriented due to a lack of information. Obstetricians and midwives have an important educational and informative role to support oocyte donation. Specific strategy of communication and valuable targeted information are needed to motivate potential donor and achieve the objectives of the program.

Assay of GCDFP-15 by ELISA: an available method for in vitro studies of functional differentiation in human breast cancer.

An enzyme-linked immunosorbent assay (ELISA) was applied to a light protein, isolated from human breast cyst fluid (BCF) termed "gross cystic disease fluid protein - 15 Kda" (GCDFP-15), a potential differentiation marker in in vitro human breast cancer studies. The detection limits of this procedure, performed in microtiter plates, were 0.5 to 250 ng/well corresponding to 10 ng/ml to 5 micrograms/ml of sample or antigen solution. Possible cross-reaction with various antigens, especially those found in culture media, were investigated. The correlation coefficient between enzymoassay and radioimmunoassay was 0.978. The results showed that quantification of GCDFP-15 by ELISA is a specific and highly sensitive method. This procedure may be of interest in in vitro studies on the functional differentiation of breast cancer cells.

Involvement of cell surface transferrin receptor in the assessment of estradiol stimulating effect on cultured breast cancer cells.

In order to find the reasons for the conflicting results depicted during the estradiol stimulations of cultured MCF7, breast cancer cells we investigated, besides cell counts, the cell surface transferrin receptor as an additional means of assessing the effect of estradiol. In this study we report results obtained using different culture conditions, i.e. short-term or long-term phenol-red withdrawn cells grown either in calf-serum supplemented media or defined media. Our results point out concurrent variations of cell counts and transferrin receptors when short-term phenol-red withdrawn cells were grown in defined media. Discrepancies were, however, observed when short-term phenol-red withdrawn cells were grown in serum-supplemented media or when long-term phenol-red withdrawn cells were grown in defined media. In both cases, only transferrin receptors account for estradiol stimulation. These results highlight the importance of transferrin receptor measurement in cultured breast cancer cell experiments and suggest cell kinetic perturbations due, in all likelihood, to serum factors or factors secreted by long-term phenol-red withdrawn cells.

New in vitro procedures for experimental studies on the development of 11-day mouse embryo forelimb buds.

A new method has been developed for culturing 11-day mouse forelimb buds in vitro. In cultures performed with conventional procedures, skeletal pieces frequently appeared distorted and reduced in size. Moreover, forelimb buds explanted from embryos younger than a stage corresponding to 50 pairs of somites developed narrow hand plates devoid of radiated autopods. By contrast, in the new procedure using media supplemented with fetal calf serum and growth factors and enhancing distal feeding with carrier implants of catgut, enlarged pads were obtained that exhibited at least 4 digital rays in buds explanted from embryos with 40-44 pairs of somites. Compared with conventional procedures, the mean value of DNA content per limb bud was twice as great with use of our improved method. The ability of limb bud cells to proliferate and differentiate when cultured either in classical or in modified conditions, and the importance of the technical procedures, are discussed in the new prospect of in vitro developmental studies.

Chondrogenesis in mouse limb buds in vitro: effects of dibutyryl cyclic AMP treatment.

We studied the effects of dibutyryl cyclic AMP (dbcAMP) on mouse limb-bud chondrogenesis at three stages of embryonic development. After 24 h of culture, limb buds with or without a covering of ectoderm were treated with 1 mM dbcAMP for 48 h and were then compared with untreated cultured limb buds. Treatment with dbcAMP enhanced cartilaginous differentiation in organ cultures of stage-17 and -19 (according to Theiler's) limb buds, although the presence of ectoderm reduced the level of dbcAMP stimulation. By stage 20, treatment with dbcAMP irreversibly inhibited cartilaginous differentiation. These results suggest that the responsiveness of mesenchymal limb-bud cells to dbcAMP is stage related. The results of histological studies as well as of analyses of DNA content and sulphated glycosaminoglycan accumulation supported the hypothesis that dbcAMP treatment induces recruitment of initially non-chondrogenic cells whose commitment explains the enhancement of cartilaginous differentiation. Limb-bud competence for chondrogenesis throughout the three developmental stages studied is also discussed.

[Chondrogenesis in mouse limb bud in vitro. I. Effect of the ectoderm]. / Chondrogenèse dans le bourgeon de membre de souris in vitro. I. Effet de l'ectoderme.

The role of the ectoderm in the chondrogenesis of mouse limb bud mesoderm was investigated in vitro at several developmental stages by analysis of the evolution of DNA content, the accumulation of sulfated glycosaminoglycans and histochemical procedures. Young limb buds or the undifferentiated apex of older buds (stages 17 and 19 of Theiler's table) from which the ectoderm had been removed with trypsin treatment initiated a large chondrogenesis but not morphogenesis. When the ectoderm was present, these limb buds showed a polarized proximal to distal outgrowth and differentiated skeletal primordia. Mesodermal cells of stage 20 limb bud apex were able to differentiate autopodial skeletons with or without the presence of the ectoderm: cartilaginous areas of the limb skeleton seem determined at this developmental stage. These results, which show the importance of the ectoderm in limb bud morphogenesis, are compared with results obtained using other methods with mouse or bird buds.