In-Yang Park

Efficient nonadhesive ex vivo expansion of early endothelial progenitor cells derived from CD34+ human cord blood fraction for effective therapeutic vascularization

Endothelial progenitor cells (EPCs) have been shown to have therapeutic potential in ischemic disease. However, the number of EPCs for cell therapy is limited. In this study, instead of the typical adherent culture method, we investigated a more efficient, clinically applicable nonadhesive expansion method for early EPCs using cord blood‐derived cells to overcome rapid cellular senescence. After a suspension culture of isolated CD34+ cells in serum‐free medium containing each cytokine combination was maintained for 9 d, the number of expanded functional EPCs was assessed by an adherent culture assay. Compared to mononuclear cells, the CD34+ fraction was superior in its expansion of functional EPCs that could differentiate into acLDL/UEA‐1+ cells without significant cellular senescence, whereas the CD34‐ fraction showed no EPC expansion. Among the cytokine combinations tested for the CD34+ fraction, a combination (SFIb) consisting of stem cell factor (SCF), FMS‐like tyrosine kinase 3 ligand, interleukin‐3, and basic fibroblast growth factor resulted in a reproducible 64‐ to 1468‐fold EPC expansion from various cord blood origins. Interestingly, the SFIb combination displayed markedly increased EPC expansion (2.43‐fold), with a higher percentage of CD34+ cells (2.17‐fold), undifferentiated blasts (2.38‐fold) and CXCR4+ cells (1.68‐fold) compared to another cytokine combination (SCF, thrombopoietin, and granulocyte colony‐stimulating factor), although the two cytokine combinations had a similar level of total mononucleated cell expansion (~10% difference). Accordingly, the cells expanded in the SFIb combination were more effective in recovery of blood flow and neovascularization in hind‐limb ischemia in vivo. Taken together, these results suggest that the nonadhesive serum‐free culture conditions of the CD34+ fraction provide an effective EPC expansion method for cell therapy, and an expansion condition leading to high percentages of CD34+ cells and blasts is likely important in EPC expansion.—O, E., Lee, B. H., Ahn, H.‐Y., Shin, J.‐C., Kim, H.‐Y., Kim, M., Park, I. Y., Park, Y.‐G., Joe, Y. A. Efficient nonadhesive ex vivo expansion of early endothelial progenitor cells derived from CD34+ human cord blood fraction for effective therapeutic vascularization. FASEB J. 25, 159–169 (2011). www.fasebj.org

The effects of oncostatin M on trophoblast cells: Influence on matrix metalloproteinases-2 and -9, and invasion activity

Oncostatin M (OSM), a cytokine of the interleukin-6 (IL-6) family, can either promote or inhibit cell growth in various normal and tumor cells and is expressed in rheumatoid arthritis, multiple sclerosis, multiple myeloma, and other inflammatory conditions. We investigated one of the possible mechanisms involved in trophoblast invasion using the human placental cell line derived from first trimester extravillous trophoblasts (HTR8SVneo): modulation of matrix metalloproteinase (MMP)-2 and -9 expression and enzymatic activity. And we addressed also the effects of exogenous OSM on the in vitro invasion activity of HTR8SVneo cells. We found that OSM enhanced the constitutive RNA and protein expressions of MMP-2 and MMP-9 in HTR8SVneo cell lines. Also, OSM treatment increased significantly the enzymatic activity of MMP-2 on gelatin zymography. The effects OSM on enzymatic activity of MMP-9 was not significant. We found that OSM increased invasion activities of HTR8SVneo cells in time-dependent and dose-dependent manners. This study suggests that OSM enhances invasion activities of extravillous trophoblasts during the first trimester through the increased enzyme activity of gelatinases, especially MMP-2.

Effect of antiviral therapy in reducing perinatal transmission of hepatitis B virus and maternal outcomes after discontinuing them

Background/Aims There have been numerous efforts to reduce mother-to-child transmission (MTCT) of hepatitis B virus (HBV) with antiviral agents during pregnancy. However, there are limited data regarding the outcomes of pregnant women after delivery. This study was performed to evaluate the efficacy of antiviral agents in preventing MTCT of HBV and maternal long-term outcomes. Methods The HBV-infected pregnant women treated with antiviral agents to prevent MTCT were retrospectively reviewed. Forty-one pregnant women who received telbivudine or tenofovir during late pregnancy (28-34 week) were analyzed. Hepatitis B virus surface antibody (HBsAb) positivity was tested in 43 infants after 7 months of birth. Eleven mothers were followed >1 year after delivery. Results The mean HBV DNA titer before antiviral therapy was 8.67 (6.60–9.49) log copies/mL, and the median age at delivery was 32 years (range, 22–40). Eleven patients were treated with tenofovir and 30 with telbivudine. The median duration was 57 days (range, 23–100), and the median HBV DNA titer at birth was 5.06 log copies/mL (range, 2.06–6.50). Antiviral treatments were associated with significant HBV DNA reduction (P<0.001). Among 43 infants (two cases of twins), HBsAb was not detected in two, subsequently confirmed to have HBV infection. Biochemical flare was observed in two of 11 mothers followed >12 months, and an antiviral agent was administered. Conclusions Antiviral treatment during late pregnancy effectively reduced MTCT. Long-term follow-up should be required in such cases. In addition, given that maternal biochemical flare occurred in 18% of mothers, re-administration of antiviral agents might be required.

P29.17: The nasolabial dimensions of facial profile at 14–39 weeks of gestation in normal Korean fetuses

Objectives: The diagnosis of polyhydramnios is easy because of improved ultrasound imaging technology. On the other hand, approximately 60 percent of cases were considered idiopathic polyhydramnios, determining the etiology can be challenging. To date, there is no published report about association between fetal prognosis and onset of polyhydramnios. We evaluated that whether the time of diagnosis of polyhydramnios is associated with etiology. Methods: All pregnant women were diagnosed in the Kochi Health Sciences Center between April 2005 and January 2010. This was a retrospective study of 2619 singleton pregnancies excluded 152 twins and 5 triplets. Polyhrdramnios was identified sonographically by amniofluid index over 25. In addition, we investigated the time of diagnosis, severity of polyhydramnios, fetal diagnosis, changes in symptoms and neonatal diagnosis was compared with fetal diagnosis. Results: Polyhydramnios was diagnosed in 18 cases (0.69%), and 14 cases (78%) of 18 neonates had one or more anomalies. Idiapathic polyhydramnios were present in only 4 cases (22%) of polyhydramnios, tended to be determined after 34 weeks gestation. All cases of fetal death or severe sequela with polyhydramnios were diagnosed at less than 30 weeks gestation. 6 cases of polyhydramnios were diagnosed after 34 weeks gestation. 2 of 3 cases with excerbation of polyhydramonios diagnosed after birth Prader-Willi syndrome and Pena-Shokeir syndrome, respectively. There was no apparent relevance to fetal prognosis and the severity of polyhydramnios. Conclusions: There is a lot of severe fetal abnormalitirs which been diagnosed by fetal diagnosis of idiopathic polyhydramnios. In the case with worsening rapidly polyhydramnios in the 3rd trimester and of early onset before 30 weeks gestation, it is necessary to consider about underlying severe congenital abnormalities by ultrasound examination can not be determined.