Stem cell therapy in utero.

In utero stem cell transplantation represents a new and still experimental therapeutic strategy for diseases related to the hematopoietic system, i.e. hemoglobinopathies, immunodeficiency diseases and metabolic disorders. To date, a total of 21 cases of transplantations using stem cells either of fetal liver or adult bone marrow origin have been reported in the literature. Success has been limited--with the exception of one case of beta-thalassemia--to four cases with immunodeficiency diseases. In this review the broad therapeutic implications as well as potentials and limitations of this technique are summarized. Furthermore, ethical considerations based on the use of fetal cells are pointed out and a prospective view concerning experimental and clinical future perspectives including the possibility for gene therapy is presented.

Laparoscopic management of heterotopic pregnancy: a review.

Laparoscopic management of ectopic pregnancies is becoming more common. Having recently successfully treated one case of heterotopic pregnancy by laparoscopy in our department, the object of the present paper is to review the literature regarding laparoscopically treated cases of heterotopic pregnancy in order to assess benefits, complications and safety of this procedure. A Medline search yielded 10 additional cases with ampullary, interstitional or cornual tubal pregnancy, respectively. All surgical procedures were uncomplicated and all in utero pregnancies progressed normally. Despite a small number of cases, lack of complications in these reports is encouraging feasibility of laparoscopic management of heterotopic pregnancy.

[The fetus as patient: intrauterine fetal stem cell transplantation]. / Der Fetus als Patient: Intrauterine fetale Stammzelltransplantation.

In utero fetal stem cell transplantation, a still experimental treatment option, represents a new therapeutic strategy with broad implications for diseases related to the hematopoietic system. This survey reviews immunological and strategical considerations as well as previously published cases. Furthermore, ethical considerations on account of the use of fetal cells are discussed and a prospective view concerning experimental and clinical future perspectives is presented.

Current status in intrauterine fetal stem cell therapy.

In utero fetal stem cell transplantation, a today experimental treatment option, represents a new therapeutic strategy with broad implications for diseases related to the hematopoietic system. The object of the present paper is to give a review of the published literature on fetal stem cell therapy with special reference to immunological and strategical considerations. Furthermore, ethical considerations on account of the use of fetal cells are pointed out and a prospective view concerning experimental and clinical future perspectives is presented.

First trimester development of the human nigrostriatal dopamine system.

The aim of the present study was to characterize the morphological and neurochemical differentiation of mesencephalic dopaminergic neurons in human embryos, derived from elective first trimester abortions. Embryonic brain tissue was taken for analysis of tyrosine hydroxylase (TH) by immunohistochemistry and Western blot, and for analysis of endogenous dopamine (A) content using HPLC-ED. TH expression was first detected at 3.5 weeks of gestational age (Carnegie stage 11) by immunohistochemical staining of the primordial sympathetic trunk along both sides of the neural tube. In sagittal sections of the intact 4.5-week-old embryo, a small, distinct population of rounded, densely packed TH-immunoreactive perikarya with short primary processes was seen in the midbrain. During the latter half of the first trimester, the number of TH-stained cells as well as the length and number of axonal processes projecting toward and into the developing neostriatum increased rapidly. At the end of the first trimester, varicose fibers could be detected in the striatal anlage. In order to verify that TH was the antigen recognized by the antibodies used for immunohistochemistry on human tissue specimens, mesencephalic tissue of 5-10 weeks gestation was analyzed by Western blot technique. A single, homogeneous band with the apparent molecular weight of approximately 60 kDa was clearly detected at 5 weeks of age. The amount of TH/mg total protein increased at least 10-fold between 5-10 weeks of gestation. For comparison, the mesencephalon and the forebrain/basal ganglia were analyzed for endogenous DA content using HPLC-ED. DA was first detected at 5.5 weeks of gestational age in both mid- and forebrain, and DA levels were found to increase exponentially from 7 to 7.5 weeks of age, reaching 4-5.5 ng DA/mesencephalon and 50-75 ng DA/g caudate nucleus-putamen anlage at the end of the first trimester. Together, morphological and biochemical data presented here constitute evidence for a very early appearance, migration, and differentiation as well as functional development of human mesencephalic dopaminergic neurons and their projections into target areas during the first trimester.

[Intrauterine fetal stem cell transplantation]. / Intrauterine fetale Stammzelltransplantation.

In utero fetal stem cell transplantation represents a new and still experimental therapeutic strategy for diseases related to the hematopoietic system. The broad therapeutic implications as well as potentials and limitations of this new technique are presented. To date, 6 cases of fetus-to-fetus transplantation have been published in the literature.

Screening of fetal stem cells for infection and cytogenetic abnormalities.

Fetal stem cell transplantation may rely on material from therapeutic abortions. It is essential that the stem cell transplant does not transmit any microorganisms that may affect the fetus and that genetically abnormal cells are avoided. To evaluate such contamination, human fetal stem cells collected February 1992 - December 1993 were analyzed for bacterial and fungal growth, and the placentas were karyotyped. Four samples of 70 were positive for different pathogens. Serological screening of 43 women during this period resulted in five seroconversions and revealed one carrier of anti-HCV. Karyotyping revealed two abnormal findings out of 72 samples. Thus, the concept of using material from therapeutic abortions is safe.

Establishment of a tissue bank for fetal stem cell transplantation.

STUDY OBJECTIVE: To analyse the yield of fetal liver tissue in first trimester abortions and to evaluate the number of nucleated cells obtained from each fetal liver during the sixth to twelfth week of gestation. DESIGN: Prospective descriptive study: LOCATION: University Hospital. MATERIAL: Women seeking abortion during a 12 month period 1992/1993. RESULTS: Out of 1271 women seeking abortion, 152 were asked whether they were willing to donate fetal tissue for fetal transplantation. Of these women, 105 (69%) accepted the proposal and underwent a modified low suction vacuum curettage. Fetal liver tissue was obtained in 61 (58%) of these procedures. The frequency at which tissue was retrieved was strongly related to gestational age and rose from 29% in week 6 to 79% in the tenth to twelfth week of gestation. The mean number of nucleated cells obtained from each fetal liver demonstrated a concomitant increase with gestational age, rising from 16 to 43 x 10(6) per liver during these weeks of gestation. Of the 61 cases in which fetal liver was obtained, four subjects were shown to be abnormal by laboratory analyses and 11 did not alter the mandatory follow-up appointment. This left 46 cases for use in the program of fetal to fetal transplantations. CONCLUSIONS: Most women seeking abortion seem to be in favor of the idea of fetal tissue donation for the treatment of other fetuses. The possibility of obtaining fetal liver tissue and the number of fetal stem cells retrieved are closely correlated to gestational age. A tissue bank appears to facilitate the operation of a fetal to fetal stem cell transplantation program.

Morphology and growth of embryonic, human dorsal root ganglion explants in long-term culture: expression of cell type-specific markers during early differentiation.

Embryonic, human spinal ganglion explants were plated at 5-12 weeks postconceptional age and cultured for 5-50 days on a semisynthetic substrate in a serum-containing culture medium without addition of antibiotics or preconditioned medium. The growth pattern in vitro was found to be age dependent. Five- to 6-week ganglia showed a characteristic semicircular growth pattern with bidirectional extension of neurites on top of a monolayer of supportive cells. Explanted 9- to 10-week ganglia showed an extensive, multidirectional neurite outgrowth with less pronounced proliferation of nonneuronal cells. Neurite extension, fasciculation, cell migration and morphology were studied immunohistochemically with antibodies to neurofilament (NF), S-100, and the Thy-1 glycoprotein. Both NF and S-100 were expressed at 5 weeks gestational age in ganglionic neurons and in proliferating Schwann cells in contact with axonal processes, respectively. NF was homogeneously distributed in both cell somata and neurites, whereas S-100 immunoreactivity showed an intense nuclear and a weaker cytoplasmic distribution in spindle-shaped, bipolar Schwann cells. This staining pattern was conserved during differentiation in long-term culture. Thy-1 was expressed on ganglionic neurites forming fascicles by the third week in culture. However, Thy-1 was never expressed until the total age of 10 weeks. In addition, Thy-1 was found on fibroblasts from the first week in culture. The distribution of Thy-1 on the cytoplasmic membrane was similar in both cell types, showing a coarsely granulated membrane staining. The temporal as well as the spatial expression of differentiation antigens in tissue sections of early embryonic spinal cord and spinal ganglia were very similar to what was observed in vitro.

Neurochemical differentiation of human bulbospinal monoaminergic neurons during the first trimester.

The neurochemical differentiation of bulbospinal noradrenergic and serotonergic neurons has been followed in first trimester human fetuses. Analysis of microdissected CNS regions revealed detectable levels of noradrenaline (NA) and serotonin (5-HT) in pons, medulla oblongata and throughout the spinal cord from 5-6 weeks of gestation. In all regions there was a pronounced increase in tissue levels of the monoamines, especially from 8-9 weeks on. 5-HT levels were lower than NA levels except for pons, where the opposite was true. With increasing fetal age, the results seemed less consistent because of considerable interindividual variations. Using immunohistochemical localization of tyrosine hydroxylase (TH), a marker for noradrenergic neurons, immature cell bodies were seen in the brain stem at the earliest stage studied, that is at 4 weeks of gestation. Several TH and 5-HT-immunoreactive (IR) cell groups were found in pons and medulla oblongata at 5 weeks. Significant structural differentiation of TH- and 5-HT-IR cell bodies was seen during the first trimester. Immunoreactive fibers began to appear at 5 weeks in the cervical spinal cord. At 6 weeks both types of fibers could be found in the white matter throughout the entire spinal cord while fibers in gray matter appeared at 9 weeks. The number of TH-IR fibers was considerably larger than the number of 5-HT-IR fibers. This is the first time the biochemical development of human bulbospinal monoaminergic neurons during the first trimester has been described. Continued investigations of the ontogenetic growth and differentiation of these human bulbospinal monoaminergic neurons will gain necessary insight into the genetically determined capacity for plasticity, potentially possible to activate later in life in response to spinal cord injury. Further, intraspinal transplantation of CNS tissue relevant to the severed spinal cord would by necessity entail selection of embryonic cell populations. Using such therapeutic strategies, detailed knowledge of the inherent capacities of the donor tissues will be crucial.

Effects of cryopreservation on subsets of fetal liver cells.

Human fetal livers from 6 to 13 weeks postconception were analysed before and after cryopreservation. The percentages of cell subsets, detected by MoAbs, did not change significantly after cryopreservation. Compared with BM, fetal liver contained significantly smaller subsets of cells identified by MoAbs, with two exceptions. Fetal liver contained a mean of 47% M5 positive cells versus 31% in BM, and there was no difference in the numbers of CD34+ cells. The colony-forming capacity was studied: 53 colonies grew from 10(5) cells from fresh fetal liver compared with 51 colonies from cryopreserved cells. For fresh BM the corresponding value was 88 per 10(5) cells. Incubation time for fetal stem cells was 17-18 days while the corresponding time for BM cells was 8-10 days.

Decreased beta-cell function in women with previous small for gestational age infants.

Insulin, proinsulin and C-peptide responses to intravenous glucose (glucose infusion test, GIT) and insulin sensitivity were measured in women who previously and for unexplained reasons gave birth to small-for-gestational-age infants (SGA, n = 10) or appropriate-for-gestational-age infants (AGA, n = 11). Insulin sensitivity was evaluated by two different methods, somatostatin-, insulin- and glucose infusion test (SIGIT) and Bergman's minimal model method applied to the frequently samples intravenous glucose tolerance test. The two groups were comparable with regard to age, parity and body mass index. The SGA group exhibited significantly (p < 0.01) lower early (0-10 min) and late (10-60 min) insulin, C-peptide and proinsulin responses during GIT than were seen in the control AGA group. Insulin sensitivity evaluated by the two techniques was increased in the SGA group, significantly so only with the minimal model method. The insulin sensitivity index (Si) according to Bergman was 10.98 +/- 2.10 in the SGA as compared to 4.36 +/- 1.18 x 10(-4)min-1 x uU-1 in the AGA group (antilogged values +/- 95% confidence intervals). Early insulin response (GIT) and Si values were inversely correlated (r = -0.48, p < 0.05).

Effect of predilatation of the uterine cervix by laminaria tent on activity of the placenta.

Serum concentrations of hCG were determined in blood samples taken 18-20 h and immediately before vacuum aspiration in 45 women in gestational weeks 7-9, admitted for legal abortion. In 35 of the women, a laminaria tent was inserted for cervical dilatation immediately after the first blood sampling. Serum hCG values decreased significantly in the women pretreated with laminaria tent, but were unchanged in the untreated women. This finding may indicate that pretreatment with a laminaria tent induces a partial placental detachment.

PIP: At the Karolinska Institute at Huddinge University Hospital in Huddinge, Sweden, serum levels of human chorionic gonadotropin (hCG) in 35 patients who had been inserted with a laminaria tent 18-20 hours prior to and immediately after vacuum aspiration were compared with those of 10 women who chose not to have laminaria tents inserted prior to vacuum aspiration. Gestational age ranged from 7 to 9 weeks. All the pregnant women were primiparous. The mean serum hCG values did not differ greatly between the cases and controls (84,260 U/1 and 75,878 U/1, respectively). Yet the mean serum hCG levels of women treated with laminaria tents to induce dilation fell considerably more than those in the control group (76,468 U/1 vs. 84,939 U/1; p.01). The concentrations fell in all treated women. Since serum hCG values normally increase during this period, the decrease indicated that laminaria tents induced partial detachment of the placenta. The results also explained the significant bleeding and/or presence of placental tissue in the cervical canal. These results suggest that laminaria tents cause partial placental detachment as well as labor contractions.

Extraovarian Brenner tumor coexisting with serous cystadenoma. Case report.

Only rare cases of extraovarian Brenner tumors have been described. This report concerns one case of extraovarian Brenner tumor detected in close association with an extraovarian benign serous cystadenoma. Coexistence of these two tumors supports the theory of a common origin either from celomic epithelium or remnants of the embryonic mesonephric system.

Evaluation of the possible interaction of the antifungal triazole SCH 39304 with oral contraceptives in normal healthy women.

The possible interaction of the newly developed triazole antifungal SCH 39304 with low-dose combined oral contraceptives (OCs) was studied in 15 healthy women. Serum levels of ovarian and adrenocortical steroids and sex hormone-binding globulin (SHBG) were analyzed in samples collected during the OC cycles immediately before, during and after the administration of SCH 39304. The drug was given as a single 400-mg oral dose on day 7 in the second cycle. SCH 39304 did not interfere with the ovulatory inhibitory action of low-dose combined OCs as judged from serum progesterone values. Administration of SCH 39304 caused minor decreases in serum SHBG, cortisol and dehydroepiandrosterone sulfate. The mechanism(s) behind these minor changes is not known.

Haematocervix after conization diagnosed by ultrasonography.

Haematocervix is an uncommon complication after conization. This report deals with 1 case that presented with clinical signs of progressive stenosis and was diagnosed by ultrasonography. Based on analysis of the 3 previously reported cases the pathogenesis of this condition is discussed.

Short-term maximal electrical stimulation--a conservative treatment of urinary incontinence.

Seventeen women with stress and 20 with motor-urge urinary incontinence were treated ambulatorily with short-term maximal electrical stimulation. The duration of the treatment was 20 min daily for 12 weeks. According to the patients' subjective evaluation, two thirds were improved. A markedly diminished leakage of urine (greater than 50% by use of pad test) was found in 6 out of 17 women with stress, and in 13 out of 20 women with motor-urge urinary incontinence. There were no significant differences observed in urodynamic parameters between the registrations performed before and after the present electrical stimulation therapy.

Influence of the maturity of the vaginal epithelium upon the absorption of vaginally administered estradiol-17 beta and progesterone in postmenopausal women.

Serum concentrations of estradiol-17 beta and its metabolites estrone (unconjugated) and total estrone (mainly estrone sulfate) and of progesterone were measured before and after application of one vaginal suppository containing 250 micrograms estradiol-17 beta and 10 mg progesterone to 10 previously unsubstituted healthy postmenopausal women with normal pretreatment serum steroid levels. Vaginal hormone cytology was performed prior to administration of the estrogen-progesterone combination. An atrophic vaginal mucosa was found in 5 and a moderate proliferation in the other 5 women. A significant association was found between proliferation and pretreatment values of estrone, but not with other steroids. There was a clear, significant association between steroid absorption kinetics and the condition of the vaginal mucosa as indicated by cytological data: significantly higher absorption was found in women with an atrophic vaginal mucosa.

Changes in fatty acid composition of cervical mucus lecithin during pregnancy.

The fatty acid composition in cervical mucus was determined in 37 pregnant and 17 nonpregnant women using gas liquid chromatography. In both groups palmitic acid (16:0) and oleic acid (18:1) were the predominant acids comprising more than half of the total amount. Compared to nonpregnant controls, pregnant women had markedly elevated levels of oleic acid (18:1), while mean levels of myristic acid (14:0) and stearic acid (18:0) were significantly lower. These pregnancy-induced changes in fatty acid pattern could not be ascribed to the increased viscosity of cervical mucus. Only minor differences in fatty acid pattern were found between women in first trimester and at term. In contrast to nonpregnant controls, a significant negative correlation between levels of myristic acid (14:0) and docosahexaenoic acid (22:6) was found in pregnant women, indicating a redistribution of these two fatty acids during pregnancy. The present study demonstrates that pregnancy alters the fatty acid composition of cervical mucus lecithin, and that the characteristic changes are present before the 10th gestational week.