Umbilical Blood Mononuclear Cells Research Articles

Perinatal Gram-Positive Bacteria Exposure Elicits Distinct Cytokine Responses In Vitro.

During pregnancy, infections caused by the gram-positive bacteria Enterococcus faecalis (E. faecalis), Streptococcus agalacticae (S. agalacticae), and Staphylococcus aureus (S. aureus) are major reasons for preterm labor, neonatal prematurity, meningitis, or sepsis. Here, we propose cytokine responses to bacterial infections by the immature perinatal immune system as central players in the pathogenesis of preterm birth and neonatal sepsis. We aimed to close the gap in knowledge about such cytokine responses by stimulating freshly isolated umbilical blood mononuclear cells (UBMC) with lysates of E. faecalis, S. agalacticae, and S. aureus collected from pregnant women in preterm labor. Bacterial lysates and, principally, S. aureus and S. agalacticae distinctly triggered most of the eleven inflammatory, anti-inflammatory, TH1/TH2 cytokines, and chemokines quantified in UBMC culture media. Chemokines depicted the most robust induction. Among them, MIP-1β was further enhanced in UBMC from female compered to male newborn infants. Due to its stability and high levels, we investigated the diagnostic value of IL-8. IL-8 was critically upregulated in cord blood of preterm neonates suffering from infections compared to gestational age-matched controls. Our results provide novel clues about perinatal immunity, underscoring a potential value of IL-8 for the timely detection of infections and suggesting that MIP-1β constitutes an early determinant of sex-specific immunity, which may contribute, e.g., to male’s vulnerability to preterm birth.

Identification of megakaryocytes as a target of advanced glycation end products in diabetic complications in bone marrow.

To define the possible effect of diabetic conditions on megakaryocytes, the long-know precursors of platelets and lately characterized modulator of hematopoietic stem quiescence-activation transition. Megakaryoblastic MEG-01 cell culture and TPO/SCF/IL-3-induced differentiation of human umbilical blood mononuclear cells toward megakaryocytes were used to test effects of glycated bovine serum albumin (BSA-AGEs). The ob/ob mice and streptozotocin-treated mice were used as models of hyperglycemia. MTT was used to measure cell proliferation, FACS for surface marker and cell cycle, and RT-qPCR for the expression of interested genes. Megakaryocytes at different stages in marrow smear were checked under microscope. When added in MEG-01 cultures at 200μg/ml, BSA-AGEs increased proliferation of cells and enhanced mRNA expression of RAGE, VEGFα and PF4 in the cells. None of cell cycle distribution, PMA-induced platelet-like particles production, expression of GATA1/NF-E2/PU-1/IL-6/OPG/PDGF in MEG-01 cells nor TPO/SCF/IL-3 induced umbilical cord blood cells differentiation into megakaryocyte was affected by BSA-AGEs. In the ob/ob diabetic mice, MKs percentages in marrow cells and platelets in peripheral blood were significantly increased compared with control mice. In streptozotocin-induced diabetic mice, however, MKs percentage in marrow cells was decreased though peripheral platelet counts were not altered. Gene expression assay showed that the change in MKs in these two diabetic conditions might be explained by the alteration of GATA1 and NF-E2 expression, respectively. Diabetic condition in animals might exert its influence on hematopoiesis via megakaryocytes-the newly identified modulator of hematopoietic stem cells in bone marrow.

Post-Traumatic Changes in the Spinal Cord in Rats after Transplantation of Mononuclear Cells from Human Umbilical Blood Modified with the vegf and fgf2 Genes

Experiments were carried out using 25 white laboratory rats. A model of dosed contusional trauma to the spinal cord at the TVIII level in rats was used to measure the areas of pathological cavities and the numbers of myelinated fibers in the outer zones of the white matter were counted after single rapid doses of human umbilical blood mononuclear cells transfected with a plasmid carrying the vegf and fgf2 genes into the injured area. Animals of the control group received the same cells in analogous conditions, but transfected with plasmid pEGFP-N2, carrying the gene for enhanced green fluorescent protein (egfp). By postadministration day 30, the total area of pathological cavities in the outer zones of the white matter on transverse sections of the spinal cord 3 mm from the trauma epicenter in the caudal direction was more than two times smaller in animals of the experimental group than in controls. The numbers of myelinated fibers in the same white matter zones at the same distance from the trauma epicenter in the caudal and rostral directions were an average of 20% greater than in controls, while at a distance of 5 mm in the rostral direction the number was 40–70% greater. Administration of the therapeutic genes vegf and fgf2 into the injured area decreased cavitation, restricted the processes of secondary degeneration and maintained the number of myelinated fibers in the damaged spinal cord.

Human umbilical blood mononuclear cell–derived mesenchymal stem cells serve as interleukin‐21 gene delivery vehicles for epithelial ovarian cancer therapy in nude mice

Ovarian cancer causes more deaths than any other cancer of the female reproductive system, and its overall cure rate remains low. The present study investigated human umbilical blood mononuclear cell (UBMC)-derived mesenchymal stem cells (UBMC-MSCs) as interleukin-21 (IL-21) gene delivery vehicles for ovarian cancer therapy in nude mice. MSCs were isolated from UBMCs and the expanded cells were phenotyped by flow cytometry. Cultured UBMCs were differentiated into osteocytes and adipocytes using appropriate media and then the UBMC-MSCs were transfected with recombinant pIRES2-IL-21-enhancement green fluorescent protein. UBMC-MSCs expressing IL-21 were named as UBMC-MSC-IL-21. Mice with A2780 ovarian cancer were treated with UBMC-MSC-IL-21 intravenously, and the therapeutic efficacy was evaluated by the tumor volume and mouse survival. To address the mechanism of UBMC-MSC-IL-21 against ovarian cancer, the expression of IL-21, natural killer glucoprotein 2 domain and major histocompatibility complex class I chain-related molecules A/B were detected in UBMC-MSC-IL-21 and in the tumor sites. Interferon-γ-secreting splenocyte numbers and natural killer cytotoxicity were significantly increased in the UBMC-MSC-IL-21-treated mice as compared with the UBMC-MSCs or the UBMC-MSC-mock plasmid-treated mice. Most notably, tumor growth was delayed and survival was prolonged in ovarian-cancer-bearing mice treated with UBMC-MSC-IL-21. Our data provide important evidence that UBMC-MSCs can serve as vehicles for IL-21 gene delivery and inhibit the established tumor.

In vitro growth of human umbilical blood mesenchymal stem cells and their differentiation into chondrocytes and osteoblasts

Conditions for culturing and differentiation of human umbilical blood mononuclear cells in vitro were studied. The growth of mesenchymal stem cells was attained in 31 of 54 (57.4%) umbilical blood samples and morphological and immunophenotypical authenticity of these cells was confirmed. Stimulatory effects of 20% AB(IV) human serum and transforming growth factor-beta (TGF-beta) on the growth of mesenchymal stem cells were demonstrated. Osteogenic cells formed in the presence of differentiation factors ascorbic acid, dexamethasone, and beta-glycerophosphate, while chondrogenic cells developed in the presence of dexamethasone, ascorbic acid, and TGF-beta. Differentiation of mesenchymal stem cells was confirmed by histochemical and molecular genetic tests.

Are human herpes viruses or measles virus associated with esophageal achalasia?

In order to test the hypothesis that esophageal achalasia may be due to neurotropic viral damage to the esophageal myenteric plexus, esophageal tissue with or without achalasia was analyzed by polymerase chain reaction for the presence of human herpes virus DNA or measles virus RNA. The DNA and RNA were extracted from the esophageal muscle of 12 patients with achalasia and six patients with upper esophageal carcinoma. Peripheral blood mononuclear cells from eight adult volunteers and two samples of umbilical blood mononuclear cells were also used as controls. PCR amplification with a pair of primers specific for herpes simplex type 1 and 2 viruses identified 92-bp fragments in nearly all specimens, including those without achalasia. Each 92-bp fragment was confirmed to be identical to a single herpes simplex virus sequence by automated DNA sequence analysis. No amplification for five other herpes viruses or measles virus was detected. Therefore, a specific viral etiology for achalasia was not identified in this study.