Animals ; Brain ; embryology ; Ear ; embryology ; Esophagus ; embryology ; Fallopian Tubes ; embryology ; Female ; Heart ; embryology ; Humans ; Kidney ; embryology ; Liver ; embryology ; Lung ; embryology ; Male ; Organogenesis ; physiology ; Penis ; embryology ; Rabbits ; Stomach ; embryology ; Vagina ; embryology

PURPOSE: Couinaud described segment IV as being equivalent to segments II and III, as the umbilical portion of the portal vein (PV), and its equal branch of segment II, originated from the transverse portion of the PV. On the contrary, Healey suggested the presence of left lateral and medial segments, on the basis of umbilical fissure. Recently, some author have claimed the branch of segment II originated from the distal portion of the ligamentum venosum (LV), and that this branch was not equal to, only a branch of, the umbilical portion. Our goal was to evaluate the surgical anatomy of the left lobe of the liver through dissecting Korean cadavers. METHODS: The number of cadavers dissected totaled 10. PV, its branches, and the LV were dissected and the length of the transverse portion measured. The distance between the origin of the transverse portion and that of the segment II branch were also measured. RESULTS: The branch of segment II originated from the distal portion of the LV in all 10 cases. The length of the transverse portion was 18.8+/-5.8 mm, and the distance between the origins of the LV and segment II branch was 7.0+/-3.1 mm. CONCLUSION: Considering the embryology of the liver, as well as the above result, the umbilical portion and segment II branch were not equal anatomic structures. The umbilical portion and LV are equal anatomic structures. The branch of segment II is only one of the branches of the umbilical portion. We think Healey's classification is more accurate for the left lobe of the liver.
Cadaver* ; Classification ; Embryology ; Hepatectomy ; Liver* ; Portal Vein

AIMTo detect the expression of a novel protein, hemangiopoietin (HAPO), in the human fetal liver at the protein level.

METHODSMonoclonal antibodies (moAbs) against HAPO were produced by traditional hybridoma technique. Their affinities were calculated from the results of non-competitive ELISA and the antibodies were purified by protein G affinity chromatography. Expression of HAPO in the liver was detected by SDS-PAGE and Western blot.

RESULTSFive strains of moAbs were screened out in total, among which three were IgG1 and the other two were IgM. Their light trains were all belonged to kappa. The relative affinities of the three IgG1 form moAbs were 3.06 x 10(9) mol/L, 6.07 x 10(8) mol/L and 1.71 x 10(10) mol/L respectively. After purification, the purity of the moAbs could reach more than 99%. HAPO expression was detected at the protein level in the human fetal liver, and the apparent molecular weight of the nature HAPO was very close to but a little higher than our recombinant one.

CONCLUSIONHAPO was expressed in the human fetal liver at the protein level.

Antibodies, Monoclonal ; Blotting, Western ; Fetus ; Humans ; Immunoblotting ; Liver ; embryology ; metabolism ; Proteoglycans ; metabolism

The milk fat globule-EGF-factor 8 protein (MFG-E8) has been identified in various tissues, where it has an important role in intercellular interactions, cellular migration, and neovascularization. Previous studies showed that MFG-E8 is expressed in different cell types under normal and pathophysiological conditions, but its expression in hematopoietic stem cells (HSCs) during hematopoiesis has not been reported. In the present study, we investigated MFG-E8 expression in multiple hematopoietic tissues at different stages of mouse embryogenesis. Using immunohistochemistry, we showed that MFG-E8 was specifically expressed in CD34+ HSCs at all hematopoietic sites, including the yolk sac, aorta-gonad-mesonephros region, placenta and fetal liver, during embryogenesis. Fluorescence-activated cell sorting and polymerase chain reaction analyses demonstrated that CD34+ cells, purified from the fetal liver, expressed additional HSC markers, c-Kit and Sca-1, and that these CD34+ cells, but not CD34- cells, highly expressed MFG-E8. We also found that MFG-E8 was not expressed in HSCs in adult mouse bone marrow, and that its expression was confined to F4/80+ macrophages. Together, this study demonstrates, for the first time, that MFG-8 is expressed in fetal HSC populations, and that MFG-E8 may have a role in embryonic hematopoiesis.
Animals ; Antigens, CD34/analysis ; Antigens, Surface/*analysis ; Bone Marrow/ultrastructure ; Female ; Hematopoietic Stem Cells/*cytology ; Liver/embryology ; Mice/*embryology ; Milk Proteins/*analysis ; Placentation ; Pregnancy

Cholestasis ; etiology ; Gestational Age ; Humans ; Hydrops Fetalis ; physiopathology ; Infant, Newborn ; Liver Function Tests ; Male ; Tachycardia, Paroxysmal ; complications ; embryology ; physiopathology ; Tachycardia, Supraventricular ; complications ; embryology ; physiopathology

OBJECTIVETo investigate the abundance of metabolic proteins in adult and fetal human livers.

METHODSAdult liver homogenate proteins, fetal liver homogenate proteins, adult liver mitochondrial proteins and fetal liver mitochondrial proteins were obtained from fetal or adult liver tissues and examined using the antibody microarrays containing 19 liver monoclonal mitochondrial antibodies. The protein expression abundances were compared among the 4 protein fractions and the pathways related to protein metabolisms were explored.

RESULTSIn adult liver mitochondria, aldehyde oxidase and carbonyl reductase were up-regulated by 2.6 and 1.7 folds, respectively, whereas corticosteroid 11-beta-dehydrogenase isozyme 1, epoxide hydrolase 1 and fibrinogen beta chain protein were down-regulated by 1.7, 1.9 and 2.2 folds, respectively, compared to those in fetal liver mitochondria. The abundance of epoxide hydrolase 1 and glutathione transferase omega-1 was significantly different between adult and fetal liver homogenate samples.

CONCLUSIONOur results demonstrate a clear difference in the expression profiles of metabolic proteins in the liver between adults and human fetuses to allow a better understanding of the occurrence and development of the metabolic proteins and the identification of markers of liver metabolism.

Adult ; Antibodies ; genetics ; metabolism ; Fetus ; metabolism ; Humans ; Liver ; embryology ; metabolism ; Mitochondria, Liver ; metabolism ; Mitochondrial Proteins ; metabolism ; Protein Array Analysis

The aberrant epigenetic reprogramme is an important cause for abnormal development of nuclear transfer embryos. The objective of this study was to investigate the CpG island methylation profiles and relative expression levels of H19 gene in different tissues of cloned goat fetus. We detected liver, placenta, kidney, lung and heart in the dead cloned goat fetus and the age-matched normal goat fetus (control) by using bisulfite sequencing and real time PCR. Results indicated that methylation levels of the fifth CpG island of H19 gene in dead cloned goat fetus was significant high compared with that in the control in placenta (70% vs 49.41%, P < 0.05), and relative expression levels of H19 gene was significant low compared with that in the control (883.3 vs 1 264.5, P < 0.05). Reversely, the methylation levels was significant low compared with that in the control in lung (63.53% vs 88.24%, P < 0.05), and relative expression levels was significant high compared with that in the control (1 003.4 vs 515.5, P < 0.05). The differences of others groups were insignificant (P > 0.05). Results showed the abnormal DNA methylation proflies of H19 gene occurred in some tissues of cloned goat fetus, which affected normal expression levels of H19 gene, indicating that aberrant DNA methylation reprogramme may be one of the important factors for the death of cloned animals.
Animals ; Cloning, Organism ; veterinary ; CpG Islands ; DNA Methylation ; Epigenesis, Genetic ; Female ; Fetus ; metabolism ; Genomic Imprinting ; Goats ; Kidney ; embryology ; metabolism ; Liver ; embryology ; metabolism ; Lung ; embryology ; metabolism ; Nuclear Transfer Techniques ; RNA, Long Noncoding ; RNA, Untranslated ; genetics ; metabolism

OBJECTIVETo induce nonparenchymal mesenchymal stem cells (NPMSCs) differentiating into functional hepatocyte-like cells in vitro, and to identify the molecular biology and functional characteristics of those hepatocyte-like cells.

METHODSHuman NPMSCs were isolated and cultured with cell culture technique. NPMSCs were induced (on 1% Matrigel as a matrix and then submitted to 2.5 mmol/L AZA pretreatment for 10-12 h), by adding HGF 10 microg/L + FGF4 10microg/L + HGM into the culture medium. The characteristics of proliferation and growth of human NPMSCs were studied with methyl thiazolyl tetrazolium (MTT). The phenotypes of NPMSCs were identified by flow cytometry, immunohistochemistry and RT-PCR. Albumin (Alb) levels in culture supernatants were determined with ELISA. Staining for glycogen of undifferentiated NPMSCs and NPMSCs derivated hepatocyte-like cells was conducted with periodic acid-Schiff (PAS) test.

RESULTSGrowth and division of adherent cells obtained from fetal livers were good and the amount of NPMSCs resourced from each fetus could be amplified to 109 cells after 10 serial subcultivations. The phenotype of NPMSCs was CD166 positive and CD34 negative. The shape of NPMSCs plated on Matrigel with FGF4 and HGF changed from long fusiform to polygonal or round on days 21-28. The rate of cell rounding was 40% and the ratio of dikaryocytes was 5%. Immunohistochemical and RT-PCR detection showed that undifferentiated NPMSCs expressed few alpha fetoprotein (AFP) and AFP mRNA, and did not express any of the liver-specific transcription factors or cytoplasmic markers. Many cells in early induction expressed GATA4, AFP and CK18 proteins and their mRNAs, and their expressions were reduced at the late induction, but the expressions of Alb, CK18, GST-and hepatocyte transcription factor HNF1increased gradually. The ratio of Alb and CK18 positive cells was 60%. Undifferentiated NPMSCs did not produce Alb. Alb production by induced NPMSCs increased in a time-dependent manner. Glycogen storage was first seen on day 14, and maximum levels were seen after day 28.

CONCLUSIONSThere are MSCs among nonparenchymal cells of fetal livers. A high ratio of hepatocyte-like cells was obtained under our induction condition. NPMSCs differentiate firstly into hepatocyte precursors, and then differentiate into mature hepatocytes and hepatocyte-like cells with positive hepatocyte markers. The induced NPMSCs have hepatocyte specific functional features.

Cell Differentiation ; Cell Separation ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Fetus ; cytology ; Hepatocytes ; cytology ; Humans ; Liver ; embryology ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo study the expression and distribution of KDR, VEGF and CD34 in yolk sac and liver of human embryo at different development stage.

METHODSYolk sacs and livers of 15 human embryos were analyzed by the immunohistochemical SP kits for the expression of KDR, VEGF and CD34.

RESULTSKDR, VEGF and CD34 were all expressed in yolk sacs and livers of the embryos. In the intermediate liver group, the grey value of KDR and VEGF were 103.8 +/- 6.1 and 96.4 +/- 6.3, respectively, stronger than that in the late liver group which were 90.4 +/- 6.0 and 87.4 +/- 6.3, respectively (P < 0.05). A positive correlation between the levels of KDR and VEGF was observed (P < 0.05).

CONCLUSIONThe expression of KDR and CD34 in yolk sac and liver of embryo suggests the presence of hemangioblast in these organs. Interaction of KDR and VEGF might relate to survival, proliferation, migration and differentiation of hemangioblasts.

Embryo, Mammalian ; metabolism ; Humans ; Immunohistochemistry ; Liver ; embryology ; metabolism ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vascular Endothelial Growth Factor Receptor-2 ; biosynthesis ; Yolk Sac ; metabolism

AIMTo establish model of differentiation of fetal liver stem cells induced by beta-ME + BHA into neural cells in vitro;

METHODSCD34+ cells from naturally aborted human fetal liver were isolated with MACS Kit, and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). After confluent more than 80%, the 5 passage cells were induced by 10(-3) mol/L beta-mercaptoethanol (beta-ME) and 2 x 10(-4) mol/L BHA for 24 hours, and washed with PBS, and then incubated in serum-free medium for 5 hours to 5 days. The characteristics of treated cells were assayed by immunocytochemistry staining analysis.

RESULTSCells treated by beta-ME+ BHA exhibited neuronal phenotype, and expressed neuronal specific proteins such as nestin, NeuN, TrnJ-1, and NF-M, which were not found in control cells. Statistic analysis showed that 81% cells were NeuN-positive, 75% cells TuJ-1-positive, 47% cells NF-M-positive, 90% cells NSE-positive.

CONCLUSIONbeta-ME + BHA can induce human fetal liver CD34+ cells to produce neuronal specific antigens and proteins in vitro and become neuronal cells. CD34+ cells from human fetal liver possess potentials of differentiation into neural cells.

Antigens, CD34 ; Butylated Hydroxyanisole ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Liver ; cytology ; embryology ; Mercaptoethanol ; pharmacology ; Neurons ; cytology