Positive effects of cryopreserved adult or fetal liver cell transplants on hypercholesterolemia and hepatic antioxidant defenses in cholesterol-fed rabbits

The liver plays a central role in lipid metabolism and the pathophysiology of many lipid disorders leads in turn to liver cell injury. Adult hepatocyte transplants provide well-recognized metabolic support, whilst hepatic stem cells may promote liver regeneration and repair, but in both cases, any clinical application would require low temperature banking of the cells. A model of dietary hypercholesterolemia was established in rabbits over 5 months, and transplants of cryopreserved adult hepatocytes (CH) and cryopreserved fetal liver cells (CFLC) were compared to Sham transplants. Cryopreservation was performed by a two-step freezing protocol using 1.5mol/l dimethyl sulfoxide (Me(2)SO). Serum contents of cholesterol lipid classes were measured during the subsequent 4 weeks, in addition to markers of serum and liver oxidative stress. Both CH and CFLC transplantation resulted in a decrease of serum lipids during the 1st week after transplantation. The effect of CH was limited to the 1st week, but CFLC provided a sustained lipid-lowering effect over the 4 weeks. The ultimate outcome of CFLC transplantation by the end of 4 weeks was more pronounced and statistically significant for both serum total cholesterol (0.15+/-0.05 versus 3.65+/-1.4mmol/l) and high-density lipoprotein-cholesterol (0.04+/-0.01 versus 0.56+/-0.06mmol/l) compared to Sham transplants (p<0.05 in both cases). CFLC transplantation also normalized hepatic tissue antioxidant defenses, namely an increase in reduced glutathione content, and enzyme activities for catalase and glutathione reductase (all significantly higher at p<0.05 than in Sham transplants) by 4 weeks.     

Immunohistochemical analyses of cell–cell interactions during hepatic organoid formation from fetal mouse liver cells cultured in vitro

Cell–cell interactions among cell types constituting the fetal liver such as hepatoblasts, stellate cells and endothelial cells lead to functional lobule development. The present study was undertaken to investigate hepatic histogenesis in the primary culture of E12.5 mouse livers, including cell–cell and cell–matrix interactions. Fetal livers were dispersed with protease treatment and cultured for 5 days. Cellular adhesion of each hepatic cell type, gene expression and extracellular matrix deposition were analyzed by immunohistochemistry and immunoblotting. Immunohistochemical analysis demonstrated that the primary culture of fetal liver cells contained at least hepatoblasts, mesenchymal cells, endothelial cells, hemopoietic cells and Kupffer cells. Although hepatoblasts, mesenchymal cells, and endothelial cells aggregated separately in the initial step, they then formed a spheroid together, adhering to the glass slide, which led to the formation of flattened hepatic organoids. Hepatoblasts more preferentially adhered to mesenchymal cells than endothelial cells. Several extracellular matrix depositions were seen in aggregates consisting of at least hepatoblasts and mesenchymal cells within 12 h, but were poor in those lacking hepatoblasts. These data show that the primary culture of fetal liver cells contains most cell types constituting fetal livers, and may be useful for studying cell–cell interactions during liver development.     

Continuous Fetal Heart Rate Monitoring and Tocodynamometry in Routine Obstetrical Care,Fetal Stress and Non-Stress Testing,and Labor Induction in the Chimpanzee (Pan troglodytes)

The use of continuous fetal heart rate (FHR) and uterine pressure monitoring in the chimpanzee (Pan troglodytes) by external Doppler transducer and tocodynamometer is described in 1) the routine obstetrical assessment of fetal well-being, 2) oxytocin challenge (stress) testing (OCT) and non-stress testing (NST) for the diagnosis of in utero fetal distress, and 3) induction of labor by intravenous oxytocin infusion, by surgical rupture of the chorio-amniotic membranes (amniotomy), or by a combination of these techniques, as an alternative to Caesarean section for clinical, managemental, or experimental purposes. FHR traces were analyzed during a total of 57 term pregnancies for three basic characteristics: baseline rate, variability, and periodic pattern. Results indicated that continuous FHR monitoring in the chimpanzee can provide a valuable tool for fetal assessment and management of labor in any attempt to reduce the unacceptably high annual incidence of perinatal and neonatal infant mortality reported in the U.S.     

胰岛素样生长因子与哺乳动物的胚胎发育

综述了胰岛素样生长因子(IGFs)在胚胎发育过程中的表达特点和对胚胎发育的作用。许多研究表明,IGFs、IGF受体、IGF结合蛋白(IGFBPs)在不同发育阶段的胚胎中具有不同的表达特点,并具有组织特异性。不论是母体来源的、胎儿自身产生的、还是外源性的IGFs都能促进细胞分化和增殖,对胚胎发育有重要作用。     

Platelet lysate: a replacement for fetal bovine serum in animal cell culture?

A new cell culture supplement, platelet lysate, was evaluated with reference to fetal bovine serum (FBS), an established industrial medium for animal cell culture. Chemical and bacteriological profiles were conducted including the presence of platelet-derived growth factor (PDGF). PDGF was detected in the platelet lysate but it was not present in FBS. The platelet lysate medium demonstrated lack of microorganisms, mycoplasma and endotoxins. The platelet lysate was investigated in culture studies (cell growth, viability and product formation) towards a number of target cells including myelomas, hybridomas, hepatocytes, fibroblasts and epithelial cells. In general the platelet lysate medium supported cell growth and maintained viabilities comparable or superior to fetal bovine serum. Productivity studies of antibodies (hybridomas) and transferrin (hepatocytes) showed similar or enhanced production in platelet-derived medium in comparison with FBS. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of normal rat serum injected into the uterus on fetal survival in the rat

We intended to use the rat model to study the effect of autoantibodies on implantation and fetal viability. However, we have since found an effect of normal rat serum on fetal resorption rate and fetal viability. The objective of this study was to determine the consistency of this effect. Wistar strain albino rats were used for injection of 150 μl normal rat serum into the lumen of uterine horn on days L₂–L₆. The other uterine horn, used as a control, was injected with 150 μl normal saline. Percent implantation, fetal resorption rate and fetal viability were determined following the intrauterine injection of normal rat serum as compared with normal saline. A significant increase in fetal resorption rate was observed following the injection of rat serum on days L₄ and L₅ (P = 0.003 and P = 0.001, respectively). A significant decrease in fetal viability was demonstrated following the injection of rat serum on day L₅ (P = 0.01). The rat can provide a suitable animal model for further studies.     

AD-MSCs and BM-MSCs Ameliorating Effects on The Metabolic and Hepato-renal Abnormalities in Type 1 Diabetic Rats

Diabetes mellitus (DM) is one of the most serious threats in the 21th century throughout the human population that needs to be addressed cautiously. Nowadays, stem cell injection is considered among the most promising protocols for DM therapy; owing to its marked tissues and organs repair capability. Therefore, our 4 weeks study was undertaken to elucidate the probable beneficial effects of two types of adult mesenchymal stem cells (MSCs) on metabolism disturbance and some tissue function defects in diabetic rats. Animals were classified into 4 groups; the control group, the diabetic group, the diabetic group received a single dose of adipose tissue-derived MSCs and the diabetic group received a single dose of bone marrow-derived MSCs. Herein, both MSCs treated groups markedly reduced hyperglycemia resulting from diabetes induction via lowering serum glucose and rising insulin and C-peptide levels, compared to the diabetic group. Moreover, the increased lipid fractions levels were reverted back to near normal values as a consequence to MSCs injection compared to the diabetic untreated rats. Furthermore, both MSCs types were found to have hepato-renal protective effects indicated through the decreased serum levels of both liver and kidney functions markers in the treated diabetic rats. Taken together, our results highlighted the therapeutic benefits of both MSCs types in alleviating metabolic anomalies and hepato-renal diabetic complications.     

Ghrelin and obestatin: different role in fetal lung development?

Ghrelin and obestatin are two proteins that originate from post-translational processing of the preproghrelin peptide. Various authors claim an opposed role of ghrelin and obestatin in several systems. Preproghrelin mRNA is significantly expressed in airway epithelium throughout lung development, predominantly during the earliest stages. The aim of this study was to evaluate the role of ghrelin and obestatin in fetal lung development in vitro. Immunohistochemistry studies were performed at different gestational ages in order to clarify the expression pattern of ghrelin, GHS-R1a, obestatin and GPR39 during fetal lung development. Fetal rat lung explants were harvested at 13.5 days post-conception (dpc) and cultured during 4 days with increasing doses of total ghrelin, acylated ghrelin, desacyl-ghrelin, ghrelin antagonist (D-Lys(3)-GHRP-6) or obestatin. Immunohistochemistry studies demonstrated that ghrelin, GHS-R1a, obestatin and GPR39 proteins were expressed in primitive rat lung epithelium throughout all studied gestational ages. Total and acylated ghrelin supplementation significantly increased the total number of peripheral airway buds, whereas desacyl-ghrelin induced no effect. Moreover, GHS-R1a antagonist significantly decreased lung branching. Finally, obestatin supplementation induced no significant effect in the measured parameters. The present study showed that ghrelin has a positive effect in fetal lung development through its GHS-R1a receptor, whereas obestatin has no effect on lung branching.