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Objective: Previously, we have found that human culture-expanded fetal lung-derived mesenchymal stem cells (MSC) promote the engraftment of umbilical cord blood (UCB)-derived CD34((+)) cells. The high frequency of MSC in fetal lung allowed us to study whether this represented a biological feature of these cells or a property that was acquired during expansion in culture., Materials and Methods: Irradiated NOD/SCID mice (n=80) were transplanted with 0.1x10(6) UCB CD34(+) cells in the presence or absence of 10(6) primary nonexpanded or culture-expanded fetal lung, liver, or BM CD45(-) cells, or with nonexpanded fetal lung liver or BM CD45(-) cells only., Results: In comparison with transplantation of UCB CD34(+) cells only, cotransplantation of UCB CD34(+) cells and primary fetal lung or BM CD45(-) cells resulted in a significantly higher level of engraftment (% hCD45(+) cells) in BM, PB, and spleen. In addition, primary mesenchymal cells derived from adult BM had a similar promoting effect. The engraftment-enhancing effect was similar to that of culture-expanded fetal lung and BM MSC. Primary mesenchymal cells, but not culture-expanded MSC, were detected in recipient mice, suggesting that the primary cells were able to home and that this capacity was lost after expansion., Conclusions: These results show that primary mesenchymal cells from fetal lung and BM promote the engraftment of UCB-derived CD34(+) cells to a similar degree as culture-expanded MSC, indicating that it reflects a biological property of primary MSC that is preserved during expansion in culture.

Background and Objectives: We previously found that human fetal lung is a rich source of mesenchymal stem cells (MSC). Here we characterize and analyze the frequency and function of MSC in other second-trimester fetal tissues., Design and Methods: Single cell suspensions of fetal bone marrow (BM), liver, lung, and spleen were made and analyzed by flow cytometry for the expression of CD90, CD105, CD166, SH3, SH4, HLA-ABC, HLA-DR, CD34 and CD45. We assessed the frequency of MSC by limiting dilution assay., Results: The frequency of MSC in BM was significantly higher than in liver, lung, and spleen (p<0.05). On primary non-expanded cells from fetal liver, lung and spleen the number of cells positive for mesenchymal markers was significantly higher within the CD34 positive population than within the CD34 negative population. The phenotype of the culture-expanded MSC was similar for all fetal tissues, i.e. CD90, CD105, CD166, SH3, SH4 and HLA-ABC positive and CD34, CD45 and HLA-DR negative. Culture-expanded cells from all tissues were able to differentiate along adipogenic and osteogenic pathways. However, adipogenic differentiation was less in MSC derived from spleen, and osteogenic differentiation was reduced in liver-derived MSC (p<0.05)., Interpretation and Conclusions: Our results indicate that culture-expanded MSC derived from second-trimester fetal tissues, although phenotypically similar, exhibit heterogeneity in differentiating potential. We speculate that these differences may be relevant for the clinical application of MSC.

Objective: To describe the annual results in all 13 Dutch in vitro fertilisation (IVF) centres in the period 1996-2000, and to look for possible differences between individual centres and years., Design: Retrospective data collection, description and analysis., Method: The results collected on the website of the Dutch Society of Obstetrics and Gynaecology (Dutch acronym: NVOG; www.nvog.nl) in the period 1996-2000 were integrated and described, with special attention to possible differences between centres and years., Results: In 1996-2000 (5 years), 63,414 IVF or ICSI treatment cycles were started in the Netherlands, and 5,884 transfers of cryopreserved embryos were performed. The number of treatment cycles increased over the years, particularly the number of ICSI cycles. The total number of ongoing pregnancies was 12,991 (20.5% per started cycle; 22.5% for ICSI and 18.3% for IVF). Particularly during the first 3 years, there was an increase in these percentages (IVF: from 16.4% (1996) to 19.2% (1998); ICSI: from 18.3% (1996) to 23.9% (1998)). There were differences between the centres in both the percentage of ongoing pregnancies per started IVF/ICSI cycle (range 13.7-25.1%) and the percentage ICSI (14-61%) and cryo-transfers per total number of treatment cycles (0-26%). It was estimated that, during this 5-year period, 1 out of every 61 Dutch neonates resulted from IVF or ICSI., Conclusion: The pregnancy-rates after IVF and ICSI increased during the study period, and were comparable with the rates in other European countries. Some important data are still missing from the inventory, for example regarding the number of embryos per transfer, multiple pregnancies, live births, congenital malformations and complications.

Suspected bowel adhesions are often considered a relative contraindication to laparoscopy. To attempt preoperative diagnoses of intraabdominal adhesions, miniculdoscopy was performed on ten patients with a miniendoscope. It was possible to establish the presence or absence of adhesions and to determine a safe abdominal trocar insertion site. The laparoscope trocar could be inserted under direct controlled vision with the culdoscope in nine cases, suggesting that this procedure averts bowel adhesions. This technique may be a contribution to safer laparoscopy.

Since 1977, routine cul-de-sac insufflation to achieve a pneumoperitoneum for laparoscopy was performed on 350 women under local anesthesia. A van Schie/van Lith intrauterine tenaculum, used together with a self-retaining Trelat speculum, guaranteed maximum stretching of the posterior vaginal fault and fixation of the anteflexed uterus necessary for an accurate 1-cm deep penetration of a pneumoperitoneum needle into the cul-de-sac. The correct position of the tip of the needle was determined precisely by means of two manometers. The failure rate in the first study of 195 procedures was 3.6% and that in the following 155 cases was 1.9%. There were no serious operative complications. The authors recommended the cul-de-sac insufflation approach for inducing pneumoperitoneum as the technique of choice.

Since 1977, cul-de-sac insufflation has been used to achieve pneumoperitoneum for laparoscopic sterilization of 570 women. The authors have previously published findings on 350 of these women (Int J Gynaecol Obstet 17(4):375, 1980), reporting technical failure rates of 3.6% and 1.9% for a series of 195 and 155 women, respectively. The failure rate for the latest series of 220 women declined to 1.4%. Reasons for this lower rate and recurring causes of the failures are given, and recommendations for the use of cul-de-sac insufflation for inducing pneumoperitoneum are suggested.