[Clinical guidelines for male infertility: controversial issues and the need to reach interdisciplinary consensus].

A review of controversial issues about the terminology on male infertility and reproductive function, which is currently used, as well as proposals for updating clinical guidelines for the diagnosis and treatment of male infertility and men in infertile couples are presented in the article. An algorithm is described, the elements of which ensure referral of patients based on the possibility and timing of restoration of reproductive function, taking into account the reproductive health of a woman, as well as increasing the likelihood of successful treatment aimed at the birth of a healthy child.

[Multipotent mesenchymal stem cells of desquamated endometrium: isolation, characterization and use as feeder layer for maintenance of human embryonic stem cell lines].

In this study, we characterize new multipotent human mesenchymal stem cell (MSC) lines derived from desquamated (shedding) endometrium in menstrual blood. The isolated endometrial MSC (eMSC) is an adhesive to plastic heterogeneous population composed mainly of endometrial glandular and stromal cells. The established cell lines meet the criteria of the International Society for Cellular Therapy for defining multipotent human MSC of any origin. The eMSCs have positive expression of CD73, CD90, CD105, CD13, CD29, CD44 markers and the absence of expression of the hematopoietic cell surface antigens CD19, CD34, CD45, CD117, CD130 and HLA-DR (class II). Multipotency of the established eMSC is confirmed by their ability to differentiate into other mesodermal cell types such as osteocytes and adipocytes. Besides, the isolated eMSC lines partially (over 50%) express the pluripotency marker SSEA-4, but do not express Oct-4. Immunofluorescent analysis of the derived cells revealed the expression of the neural precursor markers nestin and beta-III-tubulin. This suggests a neural predisposition of the established eMSC. These cells are characterized by high rate of cell proliferation (doubling time 22-23 h) and high cloning efficiency (about 60%). In vitro the eMSCs undergo more than 45 population doublings revealing normal karyotype without karyotipic abnormalilies. We demonstrate, that the mititotically inactivated eMSCs are perfect feeder cells for human embryonic stem cell lines (hESC) C612 and C910. The eMSC being a feeder culture maintain the pluripotent status of the hESC, which is revealed by the expression of Oct-4, alkaline phosphatase and SSEA-4. When co-culturing, hESC retain their morphology, proliferative rate for more than 40 passages and capability for spontaneous differentiation into embryoid bodies comprising the three embryonic germ layers. Thus, an easy and non-invasive extraction of the eMSC in menstrual blood, their multipotency and high proliferative activity in vitro without karyotypic abnormalities demonstrate the potential of use of these stem cells in regenerative medicine. Using the derived eMSCs as the feeder culture eliminates the risks associated with animal cells while transferring hESC to clinical setting.

[Peculiarities of metaphase chromosome methylation pattern in preimplantation human embryos].

Methylation pattern peculiarities revealed by immunocytochemical analysis of metaphase chromosomes from preimplanted human embryos with monoclonal antibodies against 5-methylcytosine are described. Chromosomes of 2-8-cell triploid human embryos are undermethylated, if compared to those from PHA-stimulated fetal cord blood lymphocytes. Hemimethylation (asymmetric labeling of sister chromatids) is typical for the most of embryonic chromosomes at 2-cell--blastocyst stages due most probably to a passive loss of methylation during initial cleavages. Diffuse labeling and sister chromatid exchanges are two other cytogenetic peculiarities revealed by immunofluorescent staining of early human embryos. Hypomethylation of pericentromeric heterochromatin of chromosomes 1, 9, 16 and different methylation status of some homologous chromosomes may distinguish them from metaphase chromosomes of lymphocytes. M-banding pattern typical for chromosomes from adult and cord blood lymphocytes initially appears in embryonic metaphase chromosomes as early as at a 8-cell stage to be established for most part of chromosomes of the karyotype at the morula-blastocyst stage only.

[Isolation and characterisation of continuous human embryonic stem cell lines]. / Poluchenie i kharakteristika postoiannykh linii émbrional'nykh stvolovykh kletok cheloveka.

A long-term cultivation (5-8 months) of human blastocyst-derived embryonic cells (hES) was performed. Several properties of hESs were examined to prove the state of continuous cell lines. These cells have passed through 100-175 population doublings with the average population doubling time equal to 37.0 +/- 1.5 h. Isolated hESs, referred to as HESC-1, HESC-2, HESC-3, HESC-4, cultivated on mitotically inactivated mouse embryonic fibroblasts (STO continuous cell line), formed multilayer colonies of various shape. The cells maintained stable proliferative activity, high activity of alkaline phosphatase and expression of transcription factor Oct4, and all this characterizes embryonic stem cells of different origin. Expression of hES specific cell surface antigens (SSEA-3, SSEA-4, TRA-1-81 and TRA-11-81 and TRA-1-60) was confirmed by immunofluorescence analysis with the corresponding monoclonal antibodies. An additional prove for species specificity of HESC lines is the lack of expression of mouse specific surface antigen SSEA1. The cell cycle of HESC-1 undifferentiated cells and embryoid bodies was analysed cytofluorimetrically.

[The effect of spermatozoon morphology on the fertilization rate and on disorders in early embryonic development]. / Vliianie morfologii spermatozoidov na chastotu oplodotvoreniia i narushenii rannego émbrional'nogo razvitiia.

The influence of teratozoospermia on the results of in vitro fertilization was analysed. Indices of normal sperm morphology, in cases of failed fertilization in vitro, significantly differ from those of successful fertilization. Fertilization rate in patients with less than 14.0% of normal sperm morphology was 41.9%. If the index of normal sperm morphology exceeded 14.0%, the rate of fertilization made up 62.2%. These data enable us to conclude that teratozoospermia may result in zygote development disorders and in the increased probability of embryo fragmentation.

Incidence and some perinatal problems of multiple pregnancies in a central referral hospital, Addis Ababa.

This survey covers 41,645 deliveries registered in St. Paul's Hospital, Addis Ababa (one of the 6 central referral hospitals of Ethiopia), from 1971 to 1985, with multiple deliveries comprising 2.5% of all the cases. The results obtained from analysis of these data revealed that twins occurred in the ratio of 1:41 deliveries, and triplets in 1:41(2) deliveries, this is in conformity with Hellin's statement, made in 1895. The occurrence of multiple delivery in multigravidae is shown as 1.62 times greater than that in primigravidae. There was a male predominance (52.1%) among twins. Discordant twins and triplets were observed in 31.5% and 50% respectively. In 55.9% of twin deliveries, both twins had a birthweight of less than 2500g. Head presentation of both twins occurred in 40% of the multiple births. Operative delivery was undertaken in 12.4% of all the cases.

PIP: Incidence of multiple pregnancies and the accompanying prenatal problems in a central referral hospital in Addis Ababa is explored. Examined were 41,645 deliveries from 1971-1985 at St. Paul's Hospital, Addis Ababa--1 of 6 central referral hospitals in Ethiopia. Of the 41,645 deliveries, 1,106 patients had twins, with 25 giving birth to triplets. Mean twinning incidence in 15 years was 1:40.9 ranging between 1:28 (in 1971) and 1:57 (in 1983). Annual incidence of triplets was 1:1;,665.8. Incidence of twins in the multigravidae was 43% more than in the primigravidae. Males dominated multiple births with a 52% figure, while discondant twins and triplets were observed in 31.5% respectively. Of those twins delivered, 55.9% had a birthweight less than 2500g. Head presentation of both twins occurred in 40% of the births. Operative delivery was necessary in 12.4% of the cases examined. Of particular interest to obstetricians and neonatologists, is the analysis of anthropometric data. Multiple pregnancies, and the volume of low birthweight among neonates, and perinatal mortality represent an urgency to health care professionals in Ethiopia.