Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Time course of NF- κ B activation in cocultured C 2 C 12 cells and pericytes. p65 DNA binding activity for cocultured C 2 C 12 cells and pericytes in uninjured control (CON) and scratch-injured (INJ) conditions at baseline (BSLN), 3, 6, and 24 h time points. Data are means ± SD. *Significantly increased compared to BSLN for C 2 C 12 cells. † Significantly increased compared to BSLN for pericytes.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Activation Assay, Binding Assay, Activity Assay

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Time course of MCP-1 secretion from cocultured pericytes and C 2 C 12 cells. Monocyte chemoattractant protein-1 (MCP-1) secretion by cocultured C 2 C 12 cells and pericytes in uninjured control (CON) and scratch-injured (INJ) conditions at baseline (BSLN), 3, 6, and 24 h time points. Data are means ± SD. *Significantly increased compared to BSLN for C 2 C 12 cells. † Significantly greater vs. C 2 C 12 cells at 24 h.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques:

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Pericyte transfection efficacy and efficiency in altering NF- κ B activation. (A) Representative images of pericytes transfected with vectors designed to enhance (constitutively active (c.a.) IKK β -EGFP), diminish (dominant negative (d.n.) IKK β -EGFP), or have no effect on (empty vector (e.v.) pEF6/HisB) NF- κ B activity at 24 h post transfection. (B) A luciferase reporter system was used to assess the ability of expression plasmids to alter NF- κ B expression. Data are means ± SD.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Transfection, Activation Assay, Dominant Negative Mutation, Plasmid Preparation, Activity Assay, Luciferase, Expressing

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: HMVEC proliferation is affected by pericyte NF- κ B activation in coculture. Human microvascular endothelial cell (HMVEC) number in coculture with pericytes expressing constitutively active IKK β (c.a.), dominant negative IKK β (d.n.), or empty vector control (e.v.). Data are means ± SD. *Significant difference between c.a. and d.n.

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Activation Assay, Expressing, Dominant Negative Mutation, Plasmid Preparation

Journal: Physiological Reports

Article Title: Pericyte NF- κ B activation enhances endothelial cell proliferation and proangiogenic cytokine secretion in vitro

doi: 10.14814/phy2.12309

Figure Lengend Snippet: Pericyte NF- κ B activation affects cytokine secretion in pericyte/HMVEC cocultures. Cytokine secretion of granulocyte-colony stimulating factor (G-CSF), fractalkine, interleukin 6 (IL-6), interleukin 7 (IL-7), interleukin 8 (IL-8), interferon gamma-induced protein-10 (IP-10), monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted (RANTES), and eotaxin in the cocultures of human microvascular endothelial cells (HMVECs) and pericytes that were genetically altered to express constitutively active IKK β (c.a.) or dominant negative IKK β (d.n.) in comparison to empty vector control (e.v.). Significantly greater cytokine concentration was observed in the c.a. IKK β condition compared to both e.v. and d.n. IKK β conditions for all cytokines ( P < 0.05).

Article Snippet: Human primary pericytes isolated from placental tissue were purchased from PromoCell (Heidelberg, Germany).

Techniques: Activation Assay, Dominant Negative Mutation, Plasmid Preparation, Concentration Assay

Journal: Stem Cells Translational Medicine

Article Title: Perivascular Progenitor Cells Derived From Human Embryonic Stem Cells Exhibit Functional Characteristics of Pericytes and Improve the Retinal Vasculature in a Rodent Model of Diabetic Retinopathy

doi: 10.5966/sctm.2015-0342

Figure Lengend Snippet: CD140B+CD44+ cells are derived from human embryonic stem cells (hESCs) in a matrix-dependent manner. (A): Schematic representation of matrix-dependent differentiation process for CD140B+CD44+ cells. (B): Two-color flow cytometry was used to check mesodermal specification following the differentiation schedule. All single cells were labeled with antibodies against CD140B and CD44. Obtained cells are distinguished from CD140B− and CD140B+ by the horizontal lines in each plot. Cases #1 and #2 are typical representative aspects, depending on a different starting H9-hESC line. After collagen-dependent natural selection through a simple medium change, the attached CD140B+ population showed a strong CD44 expression at Day 7 (A). Values in each quadrant plot represent percentage of population, mean ±SD (n = 6). (C, D): The change of gene context by collagen-dependent attachment step showed a big difference. Differential gene expression profiling was performed among H9-hESCs, embryoid bodies (EBs) induced by bone morphogenetic protein 4 (BMP4) (EBs) and naturally selected CD140B+CD44+ population at Day 7 (perivascular progenitor cells, PVPCs, CD140B+CD44+ population) from three independent batches. (C): Heatmap of each group. Each column represents a single microarray analysis. (D): Hierarchical clustering analysis of the global gene expression proϕiles using the average linkage and the Pearson distance. (E): Expanded CD140B+CD44+ population exhibited a unique cell morphology. Scale bar = 20 μm. (F): The cell proliferation was monitored with the CCK-8 assay at different time points. Data are means of three separated experiments ± SD. Abbreviations: BM-MSC, human bone marrow-derived mesenchyme stem cells; h, hour; hPL-PC, human placenta-derived pericyte; MEF, mouse embryonic fibroblast; OD, optical density.

Article Snippet: Both human placenta-derived pericytes (hPL-PCs) and human bone marrow-derived MSCs (BM-MSCs) were obtained from PromoCell (Heidelberg, Germany, http://www.promocell.com/ ).

Techniques: Derivative Assay, Flow Cytometry, Labeling, Selection, Expressing, Microarray, CCK-8 Assay

Journal: Stem Cells Translational Medicine

Article Title: Perivascular Progenitor Cells Derived From Human Embryonic Stem Cells Exhibit Functional Characteristics of Pericytes and Improve the Retinal Vasculature in a Rodent Model of Diabetic Retinopathy

doi: 10.5966/sctm.2015-0342

Figure Lengend Snippet: Naturally selected CD140B+CD44+ cells show the characteristics of multipotent perivascular progenitor cells (PVPCs). (A): Expanded CD140B+CD44+ cells (human embryonic stem cells derived from PVPCs; hESC-PVPCs) were labeled with the indicated antibodies. The overlaid open histogram displays immunoglobulin control against each target. Values in each quadrant plot represent percentage of population, mean ± SD (n = 8). (B): Generalized pericyte marker, NG2 (green) expressed in both hESC-PVPCs and human primary pericytes (hPL-PC). Nuclear DNA was stained with 4′,6-diamidino-2-phenylindole (blue). (C): Skeletogenic differentiation potential of hESC-PVPCs was shown in the differentiation condition of adipogenesis (left, Oil red staining) and osteogenesis (right, Alizarin red staining). (D): Smooth muscle cell (SMC) differentiation potential of hESC-PVPCs was shown in SMC differentiation media (SMDM). Quantitative polymerase chain reaction values represent mean (n = 3) ± SD (∗∗∗, p < .0001). SMC-specific markers, α-SMA (green), and CNN (red) were detected in hESC-PVPC after 6 days in SMDM differentiation. (E): Dye transfer (circle in the plot, yellow-green) increased in a coculture of DiI-labeled hESC-PVPC (PVPC-DiI, red) and Calcein-labeled other vascular lineages (upper, human umbilical vein endothelial cell, HUVEC-Calcein; lower, umbilical artery smooth muscle cell, UASMC-Calcein, green). Values in each plot represent percentage of population, mean ± SD (n = 3). Dye-transferred hESC-PVPCs were also detected on culture well (white dots). (F, G): Perivascular localization of hESC-PVPC (PVPC, DiI, red) was confirmed in three-dimensional fibrin gel bead assay with HUVEC (DiO, green). Magnified view (G) of rectangular region (F), and orthogonal projection images (white dotted lines 1 and 2). All scale bars = 100 μm. Abbreviations: α-SMA, α-smooth muscle actin; CNN, Calponin 1; d, days.

Article Snippet: Both human placenta-derived pericytes (hPL-PCs) and human bone marrow-derived MSCs (BM-MSCs) were obtained from PromoCell (Heidelberg, Germany, http://www.promocell.com/ ).

Techniques: Derivative Assay, Labeling, Marker, Staining, Real-time Polymerase Chain Reaction

Journal: American journal of reproductive immunology (New York, N.Y. : 1989)

Article Title: Placental pericytes and cytomegalovirus infectivity: implications for HCMV placental pathology and congenital disease

doi: 10.1111/aji.12728

Figure Lengend Snippet: A Placenta pericytes from Promocell cultivated in PM medium from ScienCell were allowed to develop as monolayers in chamber slides. Cell were infected with different HCMV strains: 3A-1) uninfected subconfluent growth; 3A-2) Uninfected confluent growth; 3A-3) IFA stained with alpha actin smooth muscle antibody; 3A-4) Toledo CMV infected pericytes (lab strain); 3A-5) SBCMV infected pericytes (clinical strain); 3A-6) IFA of infected pericytes expressing CMV major immediate early gene proteins; 3A-7) IFA of CMV infected pericytes expressing the pp65 and pp28 (insert) late gene proteins; and 3A-8) HCMV-GFP recombinant virus with a GFP cassette in the US28 coding region. 3B Primary placenta cytotrophoblasts isolated from term placentas cultivated in trophoblast medium from ScienCell and allowed to develop as monolayers in chamber slides. Cells were infected with different HCMV strains: 3B-1) subconfluent growth; 3B-2) confluent growth; 3B-3) IFA stained with cytokeratin-7; 3B-4) Toledo CMV infected cytotrophoblasts (lab strain); 3B-5) SBCMV infected cytotrophoblasts (clinical strain); 3B-6) IFA infected cytotrophoblasts expressing CMV major immediate early gene proteins; 3B-7) IFA of CMV infected cytotrophoblasts expressing the pp65 late gene proteins; and (3B-8) HCMV-GFP recombinant virus with a GFP cassette in the US28 coding region. 3C Primary human villous fibroblasts were cultivated in fibroblast medium from ScienCell. Cells were placed in chamber slides and infected with different HCMV strains. Mock inf = no virus; Toledo = Toledo strain of HCMV; SBCMV = clinical isolate of HCMV; US28-GFP Towne strain of HCMV with a GFP cassette in the US28 coding region; and HCMV MIE (major immediate early genes) and pp65 (late gene).

Article Snippet: All cell types supported lytic replication of a HCMV-GFP recombinant virus, as demonstrated by phase/fluorescent imaging ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 A Placenta pericytes from Promocell cultivated in PM medium from ScienCell were allowed to develop as monolayers in chamber slides.

Techniques: Infection, Staining, Expressing, Recombinant, Isolation

Journal: American journal of reproductive immunology (New York, N.Y. : 1989)

Article Title: Placental pericytes and cytomegalovirus infectivity: implications for HCMV placental pathology and congenital disease

doi: 10.1111/aji.12728

Figure Lengend Snippet: Immunohistochemical stain of placental tissue from a child with disseminated cytomegalovirus infection of the placenta. (A) cytomegalovirus infected tissue stained for the major immediate early protein (MIE) using 3,3-diaminobenzidine (DAB) colored brown. (B) Dual staining for NG2 proteoglycan for pericytes shown as dark purple and cytomegalovirus MIE stained with DAB colored brown.

Article Snippet: All cell types supported lytic replication of a HCMV-GFP recombinant virus, as demonstrated by phase/fluorescent imaging ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 A Placenta pericytes from Promocell cultivated in PM medium from ScienCell were allowed to develop as monolayers in chamber slides.

Techniques: Immunohistochemical staining, Staining, Infection

Journal: American journal of reproductive immunology (New York, N.Y. : 1989)

Article Title: Placental pericytes and cytomegalovirus infectivity: implications for HCMV placental pathology and congenital disease

doi: 10.1111/aji.12728

Figure Lengend Snippet: Luminex analysis of HCMV induction of soluble proinflammatory and angiogenic cytokines in placenta pericytes. Cytokine profiles of SBCMV-infected placental pericytes by Luminex analysis 24 h post-infection. Results from cells exposed to medium only are shown as solid black bars, cells exposed to heat-killed SBCMV are shown as shaded bars, and results from cells exposed to the SBCMV clinical isolate are shown as striped bars. Results are included for (A) MCP-1/CCL2, (B) VEGF, (C) IL-8, (D) RANTES/CCL5, (E) IL-6, (F) MMP9 and (G) TIMP-1. Results are given in picograms (pg) or nanograms per ml. Results were determined by analyzing a minimum of 20 beads collected per assay and the median result is shown.

Article Snippet: All cell types supported lytic replication of a HCMV-GFP recombinant virus, as demonstrated by phase/fluorescent imaging ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 A Placenta pericytes from Promocell cultivated in PM medium from ScienCell were allowed to develop as monolayers in chamber slides.

Techniques: Luminex, Infection

Journal: American journal of reproductive immunology (New York, N.Y. : 1989)

Article Title: Placental pericytes and cytomegalovirus infectivity: implications for HCMV placental pathology and congenital disease

doi: 10.1111/aji.12728

Figure Lengend Snippet: A. Time course analysis of HCMV-GFP infection of placenta villous cells. A graph indicating the number of infected HCMV-GFP positive villous fibroblasts (open bars), trophoblasts (gray bars), and placental pericytes (black bars) per 1.25×105 total cells over the time course 12, 24, 48 and 96 h post-infection. 5B. HCMV infected cultures of villous fibroblasts, trophoblasts, and placental pericytes at 96 h post-infection showing GFP positive cells in pericytes, villous fibroblasts, and trophoblasts 5C. (A) Phase contrast image of a tri-cell culture mixture of villous fibroblasts, trophoblasts, and pericytes representing the placental villous core. (B) A live/dead stain of the villous tri-cell culture. (C) Dual labeled IHC of the tri-cell villous culture. Villous trophoblasts stained with cytokeratin-7 antibody (brown), placental pericytes stained with CD31 antibody (red), and villous fibroblasts are unstained, color chart shown. All images have a total magnification of 200×.

Article Snippet: All cell types supported lytic replication of a HCMV-GFP recombinant virus, as demonstrated by phase/fluorescent imaging ( ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 A Placenta pericytes from Promocell cultivated in PM medium from ScienCell were allowed to develop as monolayers in chamber slides.

Techniques: Infection, Cell Culture, Staining, Labeling