Umbilical Cord Tissue as a Source of Young Cells for the Derivation of Induced Pluripotent Stem Cells Using Non-Integrating Episomal Vectors and Feeder-Free Conditions.

The clinical application of induced pluripotent stem cells (iPSC) needs to balance the use of an autologous source that would be a perfect match for the patient against any safety or efficacy issues that might arise with using cells from an older patient or donor. Drs. Takahashi and Yamanaka and the Office of Cellular and Tissue-based Products (PMDA), Japan, have had concerns over the existence of accumulated DNA mutations in the cells of older donors and the possibility of long-term negative effects. To mitigate the risk, they have chosen to partner with the Umbilical Cord (UC) banks in Japan to source allogeneic-matched donor cells. Production of iPSCs from UC blood cells (UCB) has been successful; however, reprogramming blood cells requires cell enrichment with columns or flow cytometry and specialized growth media. These requirements add to the cost of production and increase the manipulation of the cells, which complicates the regulatory approval process. Alternatively, umbilical cord tissue mesenchymal stromal cells (CT-MSCs) have the same advantage as UCB cells of being a source of young donor cells. Crucially, CT-MSCs are easier and less expensive to harvest and grow compared to UCB cells. Here, we demonstrate that CT-MSCs can be easily isolated without expensive enzymatic treatment or columns and reprogramed well using episomal vectors, which allow for the removal of the reprogramming factors after a few passages. Together the data indicates that CT-MSCs are a viable source of donor cells for the production of clinical-grade, patient matched iPSCs.

MiR-20a-containing exosomes from umbilical cord mesenchymal stem cells alleviates liver ischemia/reperfusion injury.

Mesenchymal stem cells (MSCs) have been proved to exert considerable therapeutic effects on ischemia-reperfusion (I/R)-induced injury, but the underlying mechanism remains unknown. In this study, we aimed to explore the potential molecular mechanism underlying the therapeutic effect of MSCs-derived exosome reinforced with miR-20a in reversing liver I/R injury. Quantitative real-time polymerase chain reaction, Western blot, and IHC were carried out to compare the differential expressions of miR-20a, Beclin-I, FAS, Caspase-3, mTOR and P62 in IR rats and normal rats. TUNEL was performed to assess IR-induced apoptosis in IR rats, and luciferase assay was used to confirm the inhibitory effect of miR-20a on Beclin-I and FAS expression. Among the 12 candidate microRNAs (miRNAs), miR-486, miR-25, miR-24, miR-20a,miR-466 and miR-433-3p were significantly downregulated in I/R. In particular, miR-20a, a miRNA highly expressed in umbilical cord-derived mesenchymal stem cells, was proved to bind to the 3' UTR of Beclin-I and FAS to exert an inhibitory effect on their expressions. Since Beclin-I and FAS were aberrantly upregulated in IR, exosomes separated from UC-MSCs showed therapeutic efficacy in reversing I/R induced apoptosis. In addition, exosomes reinforced with miR-20a and separated from UC-MSCs almost fully alleviated I/R injury. Furthermore, our results showed that miR-20a could alleviate the abnormal expression of genes related to apoptosis and autophagy, such as active Caspase-3, mTOR, P62, and LC3II. This study presented detailed evidence to clarify the mechanism underlying the therapeutic efficacy of UC-MSCs in the treatment of I/R injury.

Human mesenchymal stem cell sheets in xeno-free media for possible allogenic applications.

Cell-based therapies are increasingly focused on allogeneic stem cell sources because of several advantages in eliminating donor variability (e.g., aging and disease pathophysiology) affecting stem cell quality and in cell-banked sourcing of healthy donors to enable "off-the-shelf" products. However, allogeneic cell therapy is limited by host patient immunologic competence and inconsistent performance due to cell delivery methods. To address allogeneic cell therapy limitations, this study developed a new allogeneic stem cell sheet using human umbilical cord mesenchymal stem cells (hUC-MSC) that present low antigenicity (i.e., major histocompatibility complex, MHC). Optimal conditions including cell density, passage number, and culture time were examined to fabricate reliable hUC-MSC sheets. MHC II antigens correlated to alloimmune rejection were barely expressed in hUC-MSC sheets compared to other comparator MSC sheets (hBMSC and hADSC). hUC-MSC sheets easily graft spontaneously onto subcutaneous tissue in immune-deficient mice within 10 minutes of placement. No sutures are required to secure sheets to tissue because sheet extracellular matrix (ECM) actively facilitates cell-target tissue adhesion. At 10 days post-transplantation, hUC-MSC sheets remain on ectopic target tissue sites and exhibit new blood vessel formation. Furthermore, implanted hUC-MSC sheets secrete human HGF continuously to the murine target tissue. hUC-MSC sheets described here should provide new insights for improving allogenic cell-based therapies.

Impact of Mothers' Age on Telomere Length and Human Telomerase Reverse Transcriptase Expression in Human Fetal Membrane-Derived Mesenchymal Stem Cells.

Age-related cellular changes and limited replicative capacity of adult mesenchymal stem cells (MSCs) are few of the challenges confronting stem cell research. MSCs from human fetal membranes (hFM-MSCs), including placental, umbilical cord, and amniotic membrane, are considered an alternative to adult MSCs. However, the effect of mothers' age on hFM-MSC cellular properties is still not clearly established. This study aimed to evaluate the effect of mothers' age on hFM-MSC telomere length, telomerase activity, and proliferation ability in three different age groups: GI (20-29 years), GII (30-39 years), and GIII (≥40 years). hFM samples were collected from pregnant women ≤37 weeks after obtaining consent. hFM-MSCs were isolated and cultured to characterize them by flow cytometry and assess proliferation by MTT assay and doubling time. Telomere length and expression levels of human telomerase reverse transcriptase were assessed by quantitative real-time polymerase chain reaction (qRT-RCR). hFM-MSCs in the three age groups were spindle-shaped, plastic-adherent, and exhibited high proliferation rates and strong expression of hMSC markers. GI showed the longest telomere length in hMSCs in various FM regions, whereas GIII showed the highest level of telomerase expression. There was no difference in telomere length between GII and GIII, and both groups showed the same hMSC characteristics. In conclusion, although the hFM-MSCs derived from different fetal membranes maintained the MSC characteristics in all study groups, the hFM-MSCs of older mothers had shorter telomeres and higher telomerase activity and proliferation rate than did those derived from younger mothers. Thus, the hFM-MSCs of older mothers could be unsuitable for expansion in vitro or stem cell therapy. Determination of telomere length and telomerase expression level of hFM might help characterizing and understanding the biological differences of hFM-MSCs in different age groups.

In vitro toxicity screening of magnetite nanoparticles by applying mesenchymal stem cells derived from human umbilical cord lining.

Despite the growing interest in nanoparticles (NPs), their toxicity has not yet been defined and the development of new strategies and predictive models are required. Human stem cells (SCs) offer a promising and innovative cell-based model. Among SCs, mesenchymal SCs (MSCs) derived from cord lining membrane (CL) may represent a new species-specific tool for establishing efficient platforms for primary screening and toxicity/safety testing of NPs. Superparamagnetic iron oxide NPs, including magnetite (Fe3 O4 NPs), have aroused great public health and scientific concerns despite their extensive uses. In this study, CL-MSCs were characterized and applied for in vitro toxicity screening of Fe3 O4 NPs. Cytotoxicity, internalization/uptake, differentiation and proliferative capacity were evaluated after exposure to different Fe3 O4 NP concentrations. Data were compared with those obtained from bone marrow (BM)-MSCs. We observed, at early passages (P3), that: (1) cytotoxicity occurred at 10 µg/mL in CL-MSCs and 100 µg/mL in BM-MSCs (no differences in toxicity, between CL- and BM-MSCs, were observed at higher dosage, 100-300 µg/mL); (2) cell density decrease and monolayer features loss were affected at ≥50 µg/mL in CL-MSCs only; and (3) NP uptake was concentration-dependent in both MSCs. After 100 µg/mL Fe3 O4 NP exposures, the capacity of proliferation was decreased (P5-P9) in CL-MSCs without morphology alteration. Moreover, a progressive decrease of intracellular Fe3 O4 NPs was observed over culture time. Antigen surface expression and multilineage differentiation were not influenced. These findings suggest that CL-MSCs could be used as a reliable cell-based model for Fe3 O4 NP toxicity screening evaluation and support the use of this approach for improving the confidence degree on the safety of NPs to predict health outcomes.

Maternal Hematological Parameters and Placental and Umbilical Cord Histopathology in Intrauterine Growth Restriction.

OBJECTIVE: To investigate the placental and umbilical cord histopathology in intrauterine growth restriction (IUGR) and their relation to second-trimester maternal hematological parameters. MATERIALS AND METHODS: Patients were selected for the IUGR group based on estimated fetal weight below the 10th percentile. Patients were recruited into the control group randomly. Patients were followed up with ultrasound, and blood samples were taken between the 20th and 24th gestational weeks. After delivery and formalin fixation, weight and volume of the placenta were recorded and histologic samples were processed. RESULTS: Maternal platelet count strongly correlates with placental weight (r = 0.766). On the other hand, neonatal weight correlates with placental volume (r = 0.572) rather than with placental weight (r = 0.469). Umbilical arterial lumen cross-sectional area correlates with birth weight (r = 0.338). CONCLUSIONS: Maternal hematological parameters do not seem to affect neonatal outcome. Our main findings are the correlation of maternal platelet count with placental weight, the correlation of placental volume with birth weight being stronger than the correlation of placental weight with birth weight, and the correlation of umbilical artery lumen cross-sectional area with neonatal weight. Mild histopathologic alterations might occur in normal pregnancies; however, sufficient fetal nutrition can be maintained. This compensatory function of the placenta seems to be insufficient when two or more pathologies are present, which is characteristic for IUGR.

Role of VEGF in the differential growth between the fetal and placental ends of the umbilical cord.

INTRODUCTION: The umbilical cord (UC) is a vital structure; its alterations affect the newborn and neurological impact can be permanent. Paradoxically, factors that determine it remain unknown. We explore the differential VEGF protein expression in the UC's proximal and distal portions in relation to the hypothesis that the UC has differential growth and that VEGF plays a role in it. METHODS: An observational analytical study was performed. One UC segment was taken proximal to fetus and another distal; both were randomly processed; VEGF immunohistochemical analysis was performed; two blinded pathologists read results. RESULTS: Forty-eight newborns were included. Protein expression between the two edges of the umbilical cord, in any kind of cells, was interpreted. Endothelium, amnion, and stromal cells expressed VEGF; the first two were not different between opposite ends. Stromal cells had differential expression: higher in the proximal to the fetus portion. CONCLUSION: Knowledge of molecular factors is necessary. UC cells widely expressed VEGF, possibly contributing to UC growth. Even though stromal cell expression was different, the interaction with activity close to the fetus must be explored.

Differentiation of human umbilical cord mesenchymal stem cells into germ-like cells in mouse seminiferous tubules.

Our previous study demonstrated that human umbilical cord mesenchymal stem cells (HUMSCs) were capable of differentiation into germ cells in vitro. To assess this potential in vivo, HUMSCs were microinjected into the lumen of seminiferous tubules of immunocompetent mice, which were treated with busulfan to destroy endogenous spermatogenesis. Bromodeoxyuridine labeling studies demonstrated that HUMSCs survived in the tubule for at least 120 days, exhibited a round cell shape typical of proliferating or differentiating germ cells, migrated to the basement of the tubule, where proliferating spermatogonia reside and returned to the luminal compartment, where differentiating spermatids and spermatozoa reside. The migration pattern resembled that of germ cell development in vivo. Immunohistochemical and colocalization studies revealed that transplanted HUMSCs expressed the germ cell markers octamer-binding transcription factor 4, α6 integrin, C-kit and VASA, confirming the germ cell differentiation. In addition, it was observed that tubules transplanted with HUMSCs exhibited marked improvement in the histological features damaged by the chemotherapeutic busulfan, as judged by morphology and quantitative histology. Taken together, these data demonstrated the capacity of HUMSCs to form germ cells in the testes and to repair testicular tissue. These findings suggest a potential utility of HUMSCs to treat the infertility and testicular insufficiency caused by cancer therapeutics.

Secretome of mesenchymal progenitors from the umbilical cord acts as modulator of neural/glial proliferation and differentiation.

It was recently shown that the conditioned media (CM) of Human Umbilical Cord Perivascular Cells (HUCPVCs), a mesenchymal progenitor population residing within the Wharton Jelly of the umbilical cord, was able to modulate in vitro the survival and viability of different neuronal and glial cells populations. In the present work, we aimed to assess if the secretome of HUCPVCs is able to 1) induce the differentiation of human telencephalon neural precursor cells (htNPCs) in vitro, and 2) modulate neural/glial proliferation, differentiation and survival in the dentate gyrus (DG) of adult rat hippocampus. For this purpose, two separate experimental setups were performed: 1) htNPCs were incubated with HUCPVCs-CM for 5 days after which neuronal differentiation was assessed and, 2) HUCPVCs, or their respective CM, were injected into the DG of young adult rats and their effects assessed 7 days later. Results revealed that the secretome of HUCPVCs was able to increase neuronal cell differentiation in vitro; indeed, higher densities of immature (DCX(+) cells) and mature neurons (MAP-2(+) cells) were observed when htNPCs were incubated with the HUCPVCs-CM. Additionally, when HUCPVCs and their CM were injected in the DG, results revealed that both cells or CM were able to increase the endogenous proliferation (BrdU(+) cells) 7 days after injection. It was also possible to observe an increased number of newborn neurons (DCX(+) cells), upon injection of HUCPVCs or their respective CM. Finally western blot analysis revealed that after CM or HUCPVCs transplantation, there was an increase of fibroblast growth factor-2 (FGF-2) and, to a lesser extent, of nerve growth factor (NGF) in the DG tissue. Concluding, our results have shown that the transplantation of HUCPVCs or the administration of their secretome were able to potentiate neuronal survival and differentiation in vitro and in vivo.

CS5931, a novel polypeptide in Ciona savignyi, represses angiogenesis via inhibiting vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs).

CS5931 is a novel polypeptide from Ciona savignyi with anticancer activities. Previous study in our laboratory has shown that CS5931 can induce cell death via mitochondrial apoptotic pathway. In the present study, we found that the polypeptide could inhibit angiogenesis both in vitro and in vivo. CS5931 inhibited the proliferation, migration and formation of capillary-like structures of HUVECs (Human Umbilical Vein Endothelial Cell) in a dose-dependent manner. Additionally, CS5931 repressed spontaneous angiogenesis of the zebrafish vessels. Further studies showed that CS5931 also blocked vascular endothelial growth factor (VEGF) production but without any effect on its mRNA expression. Moreover, CS5931 reduced the expression of matrix metalloproteinases (MMP-2 and MMP-9) both on protein and mRNA levels in HUVEC cells. We demonstrated that CS5931 possessed strong anti-angiogenic activity both in vitro and in vivo, possible via VEGF and MMPs. This study indicates that CS5931 has the potential to be developed as a novel therapeutic agent as an inhibitor of angiogenesis for the treatment of cancer.

Umbilical cord length in singleton gestations: a Finnish population-based retrospective register study.

INTRODUCTION: Many complications of pregnancy and delivery are associated with umbilical cord length. It is important to examine the variation in length, in order to identify normal and abnormal conditions. Moreover, the factors influencing cord growth and development are not precisely known. OBJECTIVE: The main objectives were to provide updated reference charts for umbilical cord length in singleton pregnancies and to evaluate potential factors affecting cord length. METHODS: Birth register data of 47,284 singleton pregnant women delivering in Kuopio University Hospital, Finland was collected prospectively. Gender-specific centile charts for cord length from 22 to 44 gestational weeks were obtained using generalized additive models for location, scale, and shape (GAMLSS). Gestational, fetal, and maternal factors were studied for their potential influence on cord length with single variable analysis and stepwise multiple linear regression analysis. RESULTS: Cord length increased according to gestational age, while the growth decelerated post-term. Birth weight, placental weight, pregravid maternal body mass index, parity, and maternal age correlated to cord length. Gestational diabetes and previous miscarriages were associated with longer cords, while female gender and placental abruption were associated with shorter cords. DISCUSSION AND CONCLUSIONS: Girls had shorter cords throughout gestation although there was substantial variation in length in both genders. Cord length associated significantly with birth weight, placental weight, and gestational age. Significantly shorter cords were found in women with placental abruption. This important finding requires further investigation.

Utility of cryopreserved umbilical cord tissue for regenerative medicine.

MSCs can be isolated from adult sources such as bone marrow and adipose tissue. In contrast to these adult tissue sources, harvesting MSCs from cord tissue is a non-invasive procedure and poses no risk to the donor. Stem cell banks offer the opportunity to cryopreserve cord tissue as a source of MSCs for future autologous or allogeneic stem cell based regenerative medicine applications. There is little published data however, characterizing MSCs isolated from cryopreserved cord tissue. The goal of this study was to determine if MSCs isolated from cryopreserved cord tissue are functionally equivalent to MSCs isolated from fresh cord tissue. Umbilical cords were collected from 10 donors. Cords were segmented into 4-6 inch pieces and either cryopreserved or used immediately. Fresh and thawed cord segments were cultured in 7-14 days for outgrowth of MSCs. MSCs were analyzed by FACS for CD45, CD73, CD90 and CD105 expression. FACs analysis confirmed cells isolated from both fresh and frozen tissue expressed MSC markers. Adherent cells were obtained from both fresh and cryopreserved cord tissue segments at a similar plating efficiency. There was no difference in either the number or time of population doublings. MSCs isolated from fresh and frozen tissue were capable of differentiating along adipogenic, chondrogenic, osteogenic and neurogenic pathways, as confirmed by histology and RT-PCR analysis of tissue specific mRNAs. No significant functional differences were observed between MSCs from frozen cord tissue as compared to fresh cord tissue. Cryopreserving cord tissue allows for isolation of MSCs at the point of care when the specific clinical application is known. This may be advantageous as MSC isolation protocols continue to be optimized dependent on intended use.

Umbilical cord diameter percentile curves and their correlation to birth weight and placental pathology.

OBJECTIVE: The aims of this study were to develop a nomogram of umbilical cord diameter (UCD) for pathologic examination of the placenta, to identify the umbilical cord components responsible for variations in UCD, and to examine the relationship between UCD and other placental pathologic features and perinatal outcome. STUDY DESIGN: We prospectively collected 497 umbilical cords between 18 and 41 weeks' gestation over a 1-year period. Fresh-tissue UCD were grouped according to gestational age and compared to sonographic and histological measurements. Associations between UCD percentile and placental pathologic findings or obstetrical outcomes were examined. RESULTS: Mean UCD increased with gestational age until a plateau at 1.0 cm in the third trimester, a value that was 0.56 cm less than sonographic measurements prior to delivery and 0.17 cm greater than UCD measured histologically. Umbilical cord components varied with UCD percentile, with umbilical vessel area increased in thick cords (p < 0.001) and Wharton's jelly area reduced in thin cords (p = 0.002). Thin umbilical cords were associated with at least one pathologic histological placental finding (p = 0.02), low placental weight (p < 0.001), single umbilical artery (p = 0.02), marginal cord insertion (p = 0.01), and low infant birth weight (p < 0.001). CONCLUSIONS: This study provides reference curves for post-delivery UCD from 18 to 41 weeks' gestation for use by perinatal pathologists. We show that increased UCD is a function of increased umbilical blood vessel volume and decreased UCD is a function of decreased Wharton's jelly volume. UCD shows a strong association with placental and infant birth weight.

Overestimation of umbilical cord coiling index with segmental versus total length assessment.

The umbilical cord is the only communication between the fetus and the placenta and, not surprisingly, lesions or conditions affecting it may have detrimental effects in both. One important feature of the umbilical cord is its coiling index (UCI), with hypo- and hypercoiling being associated with fetal thrombotic vasculopathy, intolerance of labor, intrauterine growth restriction, cord stricture, thrombosis of cord and chorionic blood vessels, and fetal demise. It is essential that every placenta report include the UCI. The UCI could also be assessed prenatally, but there is currently no way of accurately assessing the entire length of the umbilical cord. The aim of this study was to compare UCI measured in a segment of cord 10 cm long (UCI-10) and over its total length (UCI-T). One hundred fifty consecutive placenta reports in which both measurements were recorded were retrieved from the files and analyzed. Gestational age ranged from 16 to 42 weeks, with a mean of 33.67 ± 5.96 weeks and a median of 36 weeks. Mean UCI-10 was 0.4360 ± 0.2625 coils/cm and mean UCI-T was 0.3530 ± 0.2022 coils/cm; the difference between these measurements was highly statistically significant (P < 0.0001). Counting the number of umbilical cord coils in 10 cm led to an overestimation of the UCI-T by 23.5%; it can be concluded, therefore, that the latter should be used.

First trimester umbilical cord and vitelline duct measurements using virtual reality.

BACKGROUND: the umbilical cord and vitelline duct are of vital importance to the fetus, but they are rarely the subject of first trimester two-dimensional (2D) ultrasound evaluation due to the complexity of their shape and morphology. Virtual reality (VR) allows efficient visualisation and measurement of complex structures like the umbilical cord and vitelline duct. AIM: to measure normal first trimester human growth of the umbilical cord length (UCL) and vitelline duct length (VDL) using a VR system; and to correlate both measurements with the gestational age (GA) and crown-rump length (CRL) and the VDL with the yolk sac volume (YSV). STUDY DESIGN: prospective cohort study. Serial three-dimensional (3D) ultrasound measurements were performed from six to 14weeks GA, resulting in 125 3D volumes. These volumes were analysed using an I-Space VR system. SUBJECTS: Thirty-two healthy pregnant women with an ongoing, normal pregnancy. OUTCOME MEASURES: the UCL, VDL, YSV and other related structures were measured. RESULTS: The UCL, measurable in 55% of cases, was positively correlated to advancing GA and CRL (p<0.001). The VDL could be measured in 42% of cases and showed a positive relationship with GA and CRL (p<0.001). There was a significant (p<0.001) relationship between YSV and VDL. CONCLUSIONS: the present study, facilitated by a VR system, is the first to provide an in-vivo longitudinal description of normal first trimester growth of the human umbilical cord and vitelline duct. Further studies will reveal whether these parameters can be used in detection of abnormal fetal development.

The change of umbilical cord components in intrauterine growth restriction comparative with normal growth fetuses by using sonographic measurement.

OBJECTIVE: To determine the diameter changes of umbilical cord components in intrauterine growth restriction (IUGR) fetuses comparative with normal growth fetuses by using the ultrasonogram. MATERIAL AND METHOD: A cross sectional study was performed with 140 singleton pregnant women who was attended at Maternal-Fetal Medicine unit, Thammasat University Hospital between June, 2007 to May, 2009. The fetuses were between the gestational ages of 24 to 37 weeks at the time of data collection. Seventy pregnant women with IUGR fetuses and 70 pregnant women with normal growth (Appropriate for Gestational Age, AGA) fetuses were included. The sonogram of the umbilical cord, umbilical artery and umbilical vein diameter and circumference were obtained at the free loop of cord. Fetal weights were estimated by calculation in all cases. IUGR was defined as a fetus having an estimated fetal weight below the 10th percentile for the gestational age at time of the sonographic measurements. RESULTS: The mean age of the patients were 27.03 and 31.86 years in IUGR group and AGA group, respectively. The mean birth weight of the fetuses was 2153.60 +/- 386.13 gm and 3118.16 +/- 353.28 gm in IUGR fetuses and AGA fetuses, respectively. The result demonstrated the expected progressive increase of the umbilical cord circumference and diameter as a function of gestational age in AGA fetuses. These changes were not observed in the umbilical cord of IUGR fetuses. It was a contradictory finding that the measurement values from umbilical artery and umbilical vein IUGR fetuses were neither consistent nor correlated with fetal age. CONCLUSION: The ultrasonogram of the umbilical cord component demonstrated the increasing of umbilical cord circumference and diameter along with an increasing of gestational age in the AGA fetuses. These findings might be useful for further studies, such as early screening or prediction of adverse pregnancy outcome for high risk women.

Comparative proteomic analysis of malformed umbilical cords from somatic cell nuclear transfer-derived piglets: implications for early postnatal death.

BACKGROUND: Somatic cell nuclear transfer (scNT)-derived piglets have high rates of mortality, including stillbirth and postnatal death. Here, we examined severe malformed umbilical cords (MUC), as well as other organs, from nine scNT-derived term piglets. RESULTS: Microscopic analysis revealed complete occlusive thrombi and the absence of columnar epithelial layers in MUC (scNT-MUC) derived from scNT piglets. scNT-MUC had significantly lower expression levels of platelet endothelial cell adhesion molecule-1 (PECAM-1) and angiogenesis-related genes than umbilical cords of normal scNT piglets (scNT-N) that survived into adulthood. Endothelial cells derived from scNT-MUC migrated and formed tubules more slowly than endothelial cells from control umbilical cords or scNT-N. Proteomic analysis of scNT-MUC revealed significant down-regulation of proteins involved in the prevention of oxidative stress and the regulation of glycolysis and cell motility, while molecules involved in apoptosis were significantly up-regulated. Histomorphometric analysis revealed severe calcification in the kidneys and placenta, peliosis in the liver sinusoidal space, abnormal stromal cell proliferation in the lungs, and tubular degeneration in the kidneys in scNT piglets with MUC. Increased levels of apoptosis were also detected in organs derived from all scNT piglets with MUC. CONCLUSION: These results suggest that MUC contribute to fetal malformations, preterm birth and low birth weight due to underlying molecular defects that result in hypoplastic umbilical arteries and/or placental insufficiency. The results of the current study demonstrate the effects of MUC on fetal growth and organ development in scNT-derived pigs, and provide important insight into the molecular mechanisms underlying angiogenesis during umbilical cord development.

Valores hematologicos en recien nacidos sanos habitantes de altura: 3600 msnm / Haematologic values in normal newborns at 3600 meters above sea level

Pregunta de investigación. ¿Cuáles son los valores normales hematológicos de sangre de cordón umbilical de recien nacidos sanos de altura habitantes a 3600 m.s.n.m. cuyas madres son atendidas en el Hospital de la mujer de la ciudad de La Paz durante la gestión 2002?. Objetivo general. Conocer los valores normales hematológicos de sangre de cordón umbilical (serie roja y serie blanca) de los recien nacidos sanos habitantes de 3600 msnm antendidos en el hospital de la Mujer durante la gestión 2002. Diseño. Para responder la pregunta de investigación y cumplir con los objetivos, se diseño un estudio de corte transversal. Contexto o lugar. el estudio fue realizado en un centro de referencia como es el Hospital de la Mujer y se procesaron las muestras en el laboratorio del Instituto de Genética. Pacientes. Recien nacidos atendidos en el Hospital de la Mujer, según tamaño de muestra calculado tomando encuesta un nivel de confianza de 95 porciento, poder del 80 porciento, factor de revalencia para recien nacidos sanos de 65 porciento, dando un total de 297 pacientes; estos niños fieron ejegidos najo los criterios de inclusión y exclusión descritos. Intervención. Los criterios de inclusión fueron los RN sanos de altura (3600 m.s.n.m.) de ambos sexos. Los criterios de exclusión están referidos a Hipertensión materna, ruptura de membranas fetales por mas de 24 horas, fiebre materna, trabajo de arto mayor de 18 horas, presentaciones anormales. apgar menor a 5, etc. Una vez seleccionados los pacientes, se realizó la toma de muestra de angre de cordón umbilical (lado del niño) a libre caida, en el momento del nacimiento, vetiendo una cantidad de 3 ml. Aproximadamente en un tubo colector que contiene EDTA En laboratorio se mide la serie roja a tavés del hematócrito, hemoglobina y características morfológicas de klos eritrocitos en una placa de tinción utilizando la tinción de May Grunwald-Giensa, que al mismo tiempo nos permite analizar la fórmula Leucocitaria y os agregados plaquetarios. Se realizó el recuento de glóbulos blancos y plaquetas con la técnica UnoPett. Mediciones. Todo el equipo humano que realizó la recolección y el procesamiento de las muestras fue entrenado previamente. Se llenó un cuestionario de antecedentes generales para la identificación del niño recien nacido. En laboratorio se utilizó técnicas y equipos validados para realziar el Hto., Hb y el recuento de las células blancas según normas internacionales.

TGF-beta 3 expression during umbilical cord development and its alteration in pre-eclampsia.

Members of the TGF-beta family have been shown to play an important role in numerous tissues during development. In the present study we have investigated the spatial and temporal expression of TGF-beta 3, in human umbilical cord development. Total TGF-beta 3 protein content, assessed by immunoblotting, increased with advancing gestation as did immunostaining and mRNA in Wharton's jelly fibroblasts. Immunohistochemical analysis revealed that TGF-beta 3 was present in all cell types. Temporal changes in TGF-beta 3 expression were observed in the vascular smooth muscle cells, such that with advancing gestation TGF-beta 3 protein expression and became mostly restricted to the extracellular compartment of the vascular media. This was associated with a decrease in TGF-beta 3 mRNA expression in umbilical vascular smooth muscle cells. Of clinical significance, umbilical cords from pregnancies complicated by pre-eclampsia, showed a significant reduction in total TGF-beta 3 protein expression when compared to those of age-matched patients. Both TGF-beta 3 mRNA and protein expression were downregulated in the endothelium and smooth muscle layers of the umbilical arteries, as well as in the Wharton jelly fibroblasts. Our data demonstrate that during umbilical cord development TGF-beta 3 expression is spatially and temporally regulated and that TGF-beta 3 expression is altered in umbilical cords of pregnancies complicated by pre-eclampsia. We speculate that the downregulation of TGF-beta 3 expression found in pre-eclamptic umbilical cord may contribute to the abnormal structure and mechanical properties seen in these pathological umbilical cords.

Maternal uniparental disomy of chromosome 16 and body stalk anomaly.

We report on a fetus with placental trisomy 16, maternal uniparental disomy (UPD), and body stalk anomaly. Body stalk anomaly is a rare, fatal developmental abnormality consisting of a defective abdominal wall with abdominal organs in a sac outside the abdominal cavity covered by amnion adherent to the placenta with absence or severe shortness of the umbilical cord. Trisomy 16 was identified in the placenta in all cells. Amniocentesis was karyotypically normal. Parental origin studies showed maternal UPD for chromosome 16 in post-termination fetal tissue. The cause of the body stalk anomaly is not clearly defined. There are no other reports of placental karyotype or UPD investigations with body stalk anomaly. To our knowledge, this is the first report of placental trisomy 16, UPD in fetus, and body stalk anomaly, suggesting placental insufficiency or imprinting effects as cause of this anomaly. Am. J. Med. Genet. 94:284-286, 2000.