scholarly journals Substrate elasticity controls cell proliferation, surface marker expression and motile phenotype in amniotic fluid-derived stem cells

2013 ◽  
Vol 17 ◽  
pp. 307-316 ◽  
Author(s):  
Aleksander Skardal ◽  
David Mack ◽  
Anthony Atala ◽  
Shay Soker
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Amniotic Fluid ◽  
Surface Marker ◽  
Surface Marker Expression ◽  
Marker Expression ◽  
Substrate Elasticity

scholarly journals Shared Cell Surface Marker Expression in Mesenchymal Stem Cells and Adult Sarcomas

2012 ◽  
Vol 2 (1) ◽  
pp. 53-60 ◽  
Author(s):  
Stefan Wirths ◽  
Elke Malenke ◽  
Torsten Kluba ◽  
Simone Rieger ◽  
Martin R. Müller ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Surface ◽  
Surface Marker ◽  
Cell Surface Marker ◽  
Surface Marker Expression ◽  
Marker Expression ◽  
Cell Surface Marker Expression

Different protein of Echinococcus granulosus stimulates dendritic induced immune response

Parasitology ◽  
2015 ◽  
Vol 142 (7) ◽  
pp. 879-889 ◽  
Author(s):  
YANA WANG ◽  
QIANG WANG ◽  
SHIYU LV ◽  
SHENGXIANG ZHANG
Keyword(s):  
Immune Response ◽  
Cell Proliferation ◽  
T Cell ◽  
Echinococcus Granulosus ◽  
Surface Marker ◽  
T Cell Proliferation ◽  
Surface Marker Expression ◽  
Marker Expression ◽  
Dc Maturation

SUMMARYCystic echinococcosis is a chronic infectious disease that results from a host/parasite interaction. Vaccination with ferritin derived from Echinococcus granulosus is a potential preventative treatment. To understand whether ferritin is capable of inducing a host immune response, we investigated the response of dendritic cells (DCs) to both recombinant ferritin protein and the hydatid fluid (HF) of E. granulosus. We evaluated the immunomodulatory potential of these antigens by performing, immunocytochemistry, electron microscopy and in vivo imaging of monocyte-derived murine DCs. During antigen stimulation of DCs, ferritin cause DCs maturation and induced higher levels of surface marker expression and activated T-cell proliferation and migration. On contrary, HF failed to induce surface marker expression and to stimulate T-cell proliferation. In response to HF, DCs produced interleukin-6 (IL-6), but no IL-12 and IL-10. DCs stimulated with ferritin produced high levels of cytokines. Overall, HF appears to induce host immunosuppression in order to ensure parasite survival via inhibits DC maturation and promotes Th2-dependent secretion of cytokines. Although ferritin also promoted DC maturation and cytokine release, it also activates CD4+T-cell proliferation, but regard of the mechanism of the Eg.ferritin induce host to eradicate E. granulosus were not clear.

scholarly journals Comparison of Canine and Feline Adipose-Derived Mesenchymal Stem Cells/Medicinal Signaling Cells With Regard to Cell Surface Marker Expression, Viability, Proliferation, and Differentiation Potential

2021 ◽  
Vol 7 ◽  
Author(s):  
Metka Voga ◽  
Valerija Kovač ◽  
Gregor Majdic
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Surface Marker ◽  
Growth Media ◽  
Differentiation Potential ◽  
Surface Marker Expression ◽  
Proliferation And Differentiation ◽  
Marker Expression ◽  
In Cells

Remarkable immunomodulatory abilities of mesenchymal stem cells, also called multipotent mesenchymal stromal cells or medicinal signaling cells (MSCs), have entailed significant advances in veterinary regenerative medicine in recent years. Despite positive outcomes from MSC therapies in various diseases in dogs and cats, differences in MSC characteristics between small animal veterinary patients are not well-known. We performed a comparative study of cells' surface marker expression, viability, proliferation, and differentiation capacity of adipose-derived MSCs (ADMSCs) from dogs and domestic cats. The same growth media and methods were used to isolate, characterize, and culture canine and feline ADMSCs. Adipose tissue was collected from 11 dogs and 8 cats of both sexes. The expression of surface markers CD44, CD90, and CD34 was detected by flow cytometry. Viability at passage 3 was measured with the hemocytometer and compared to the viability measured by flow cytometry after 1 day of handling. The proliferation potential of MSCs was measured by calculating cell doubling and cell doubling time from second to eighth passage. Differentiation potential was determined at early and late passages by inducing cells toward adipogenic, osteogenic, and chondrogenic differentiation using commercial media. Our study shows that the percentage of CD44+CD90+ and CD34−/− cells is higher in cells from dogs than in cells from cats. The viability of cells measured by two different methods at passage 3 differed between the species, and finally, canine ADMSCs possess greater proliferation and differentiation potential in comparison to the feline ADMSCs.

A High-Throughput Screen to Identify Compounds Preserving Primary Human AML Stem Cells Ex-Vivo,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3587-3587
Author(s):  
Caroline Pabst ◽  
Josee Hebert ◽  
Guy Sauvageau
Keyword(s):  
Stem Cells ◽  
High Throughput ◽  
Conflicts Of Interest ◽  
Ex Vivo ◽  
Surface Marker ◽  
Leukemic Cells ◽  
High Throughput Screen ◽  
Self Renewal ◽  
Surface Marker Expression ◽  
Marker Expression

Abstract Abstract 3587 Acute myeloid leukemia (AML) is the second most frequent leukemia in adults with still high relapse rates supposed to be due to a rare population of leukemia stem cells (LSC) within the leukemic bulk not responding to conventional anti-cancer drugs. Engraftment of human leukemic cells in immunocompromised mice mimicking the human disease is considered to prove the existence of stem cells in the sample; however, the engraftment potential of primary human AML cells is usually quickly lost upon ex-vivo culture indicating that LSC have undergone differentiation or apoptosis and thus lost their stem cell properties. This lack of appropriate culture conditions enabling ex-vivo maintenance of primary human LSC is a major obstacle in the development of LSC targeted therapies. Our aim is, therefore, to identify conditions which allow the maintenance and expansion of these rare cells ex-vivo to understand their unique properties and to further elucidate cellular pathways underlying the self-renewal program of human primary LSC. We established a flow cytometry based high-throughput screen in which we tested more than 6,000 selected compounds with regards to their potential to expand primary human AML cells while preserving their phenotypic and functional characteristics. We considered alteration of surface marker expression, i.e. loss of CD34 and acquisition of markers of myeloid maturation initially not expressed on the cells, as indicator of differentiation implicating at the same time a loss of stem cells in the culture. We hypothesized that compounds that would be able to prevent these alterations in surface marker expression could have the potential to promote self-renewal of primary human LSC ex vivo. A normal-karyotype AML representing the most frequent cytogenetic group of AML in adults with FAB M1 morphology was chosen to perform the screen. Moreover, the initial CD34+CD15- phenotype allowed us to monitor changes in surface marker expression. Cells were plated in 384 well plates with a density of 5,000 cells per well. 6,160 compounds were tested with and without an inhibitor to aryl hydrocarbon receptor (AhR) and the vehicle DMSO was used as negative control resulting in a final DMSO concentration of ∼0.1% in each well, which had been proven to be not toxic to the cells. After 5 days of incubation the cells were analyzed on a BD LSRII high-throughput flow cytometer with regards to CD34 and CD15 surface expression. Compounds showing an increase of ≥45% of CD34+ and CD34+CD15- cells over DMSO compared to the AhR antagonist were selected for further evaluation. In a secondary screen we retested 32 compounds in 5 different concentrations ranging from 1:34 to 3x based on the initial screen concentration after 5 and 9 days of incubation and confirmed an increase ≥50% of relative and absolute CD34+CD15- leukemic cells after 5 days for 13 compounds and after 9 days for 7 compounds. We have initiated functional in-vitro and in-vivo experiments to validate these results and to prove the existence of LSC in cultures where compounds have been added. The ability of the newly identified compounds to preserve human LSC ex-vivo will be further evaluated using a collection of primary AML samples comprising different morphological and cytogenetic groups. Together, our observations document the feasibility of finding anti-differentiation (pro-self-renewal?) compounds of primary human AML cells that can be expanded ex-vivo. These studies represent a first key step in the identification of specific LSC targeting compounds. Disclosures: No relevant conflicts of interest to declare.

Mass Cytometric Analysis of AML Stem and Early Progenitor Cells Reveals Karyotype and Genotype-Specific Immunophenotypes That May Represent Targets for Antibody-Directed Therapies

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2380-2380
Author(s):  
Gregory K. Behbehani ◽  
Wendy J. Fantl ◽  
Bruno C Medeiros ◽  
Garry P. Nolan
Keyword(s):  
Stem Cells ◽  
Progenitor Cell ◽  
Expression Patterns ◽  
Fold Increase ◽  
Surface Marker ◽  
Leukemic Cells ◽  
Therapeutic Antibodies ◽  
Hematopoietic Stem ◽  
Surface Marker Expression ◽  
Marker Expression

Abstract Introduction: Leukemic stem cells (LSCs) are recognized as important mediators of leukemia relapse. Thus, therapeutic antibodies are in development to target antigens present on these cells. The success of this approach relies on a detailed understanding of the surface marker expression patterns of LSCs. To address this, we applied multi-parametric single-cell mass cytometry (MCM) to deeply profile the surface marker expression of all major immunophenotypic populations in bone marrow aspirates (BM) from patients with acute myeloid leukemia (AML). Methods: BMs from 41 leukemia patients (30 AML, 4 APL, 2 high-risk MDS, 5 AML in CR) and 5 healthy donors and were processed immediately after aspiration (<1min) and stored for pooled analysis with two overlapping 39-antibody MCM panels (50 markers total). All samples were barcoded, such that 20 samples (leukemia and healthy) could be combined into a single tube for simultaneous antibody staining and analysis, resulting in high precision (coefficient of variation = 10-20%). Results: Distinct AML subtype-specific patterns of cell frequencies across immunophenotypic populations were detected. Patients with core binding factor (CBF) AML (n=5) and those with adverse-risk karyotpic abnormalities (n=6) exhibited the greatest expansion of immunophenotypic hematopoietic stem cells (HSCs) and early progenitors (MPP, CMP), while patients with APL (n=4) and FLT3-ITD normal karyotype AML (NK-AML; n=11) exhibited reduced expansion of early progenitors and expansion in more mature myeloid progenitors (GMP, myelo-monoblasts). As a result of barcoding, high resolution (2-3 fold changes) measurement of surface marker expression detected multiple aberrancies across almost all identifiable immunophenotypic populations. Specifically, several genotype- and karyotype-specific trends in aberrant marker expression were observed in hematopoietic stem and progenitor cell populations (HSPCs; CD34+CD38low). FLT3-ITD samples were characterized by increased CD7, CD33, CD123, CD45, CD321, and CD99, as well as decreased CD34, CD117, and CD38. FLT3wt NK-AML samples (n=6) were characterized by increased CD99 as well as decreased CD71, CD47, CD34, and CD45. Adverse-risk karyotype samples (n=6) were characterized by increased CD99 and decreased CD47. All p values were significant (ranging from 0.02 to 4.5x10-7). While all 36 samples (AML or APL) displayed immunophenotypic abnormalities within the CD34+CD38low gated population (3.6 abnormalities on average), there were 9 samples in which an unambiguous separation into normal and leukemic populations was observed among the cells in this gate. Interestingly, among these samples, some markers were aberrantly high in leukemic cells of one subtype, and aberrantly low in leukemic cells of another, e.g. HLA-DR was extremely high in HSPCs of sample #22 (MLL rearrangement), and aberrantly low in all 4 APL samples. CD99 was most consistently elevated in the AML samples (28/36), but was normal in 6 samples, including a majority of those with t(8;21). These results suggest that therapeutic antibodies directed against molecules such as CD33 may be less effective for AML subtypes such as FLT3wt NK-AML (~40% decrease in CD33), and more effective for other subtypes such as FLT3-ITD NK-AML (7-fold increase in CD33). HSPCs from patients with FLT3-ITD mutations also displayed a 20-fold increase in CD123, which could also be therapeutically targeted. These findings, combined with our recent observation that HSPCs from FLT3-ITD NK-AML patients have an extremely low S-phase fraction, may provide a mechanistic basis for the improved disease-free survival recently reported for FLT3-ITD NK-AML patients treated with fractionated gemtuzumab ozogamicin in combination with standard therapy. Conclusions: We have developed an innovative MCM approach for the analysis of hematologic malignancies and demonstrated that different genotypic and karyotypic subsets of AML are highly varied in their immunophenotypic properties, particularly within the stem and progenitor cell compartment. These data also suggest that antigens that distinguish AML stem cells from normal stems cells are likely to be karyotype and genotype-specific. These findings have important implications for the design of therapeutic strategies in AML. Figure 1 Figure 1. Figure 2 Figure 2. Figure 3 Figure 3. Disclosures Behbehani: Fluidigm: Consultancy. Medeiros:Agios: Consulting - Ad board Other. Nolan:Fluidigm, Inc: Consultancy, Equity Ownership.

scholarly journals Jaw Periosteum-Derived Mesenchymal Stem Cells Regulate THP-1-Derived Macrophage Polarization

2021 ◽  
Vol 22 (9) ◽  
pp. 4310
Author(s):  
Fang He ◽  
Felix Umrath ◽  
Siegmar Reinert ◽  
Dorothea Alexander
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Macrophage Polarization ◽  
Polarization State ◽  
Osteonecrosis Of The Jaw ◽  
Surface Marker ◽  
Surface Marker Expression ◽  
Periosteal Cells ◽  
Immunomodulatory Properties ◽  
Marker Expression

Mesenchymal stem cells from bone marrow have powerful immunomodulatory capabilities. The interactions between jaw periosteal cells (JPCs) and macrophages are not only relevant for the application of JPCs in regenerative medicine, but this understanding could also help treating diseases like osteonecrosis of the jaw. In previous studies, we analyzed, for the first time, immunomodulatory features of 2D- and 3D-cultured JPCs. In the present work, the effects of JPCs on the polarization state of macrophages in contact coculture were analyzed. To improve the macrophage polarization study, different concentrations of PMA (5 nM, 25 nM, and 150 nM) or different medium supplementations (10% FBS, 10% hPL and 5% hPL) were compared. Further, in order to analyze the effects of JPCs on macrophage polarization, JPCs and PMA-stimulated THP-1 cells were cocultured under LPS/IFN-γ or IL-4/IL-13 stimulatory conditions. Surface marker expression of M1 and M2 macrophages were analyzed under the different culture supplementations in order to investigate the immunomodulatory properties of JPCs. Our results showed that 5 nM PMA can conduct an effective macrophage polarization. The analyses of morphological parameters and surface marker expression showed more distinct M1/M2 phenotypes over FBS supplementation when using 5% hPL during macrophage polarization. In the coculture, immunomodulatory properties of JPCs improved significantly under 5% hPL supplementation compared to other supplementations. We concluded that, under the culture condition with 5% hPL, JPCs were able to effectively induce THP-1-derived macrophage polarization.

scholarly journals Mesenchymal Stem/Stromal Cells Derived from Induced Pluripotent Stem Cells Support CD34posHematopoietic Stem Cell Propagation and Suppress Inflammatory Reaction

2015 ◽  
Vol 2015 ◽  
pp. 1-14 ◽  
Author(s):  
Mohsen Moslem ◽  
Irina Eberle ◽  
Iuliia Weber ◽  
Reinhard Henschler ◽  
Tobias Cantz
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Induced Pluripotent Stem Cells ◽  
Stromal Cells ◽  
Pluripotent Stem Cells ◽  
Surface Marker ◽  
Surface Marker Expression ◽  
Cell Source ◽  
Marker Expression ◽  
Induced Pluripotent

Mesenchymal stem/stromal cells (MSCs) represent a promising cell source for research and therapeutic applications, but their restrictedex vivopropagation capabilities limit putative applications. Substantial self-renewing of stem cells can be achieved by reprogramming cells into induced pluripotent stem cells (iPSCs) that can be easily expanded as undifferentiated cells even in mass culture. Here, we investigated a differentiation protocol enabling the generation and selection of human iPSC-derived MSCs exhibiting relevant surface marker expression profiles (CD105 and CD73) and functional characteristics. We generated such iPSC-MSCs from fibroblasts and bone marrow MSCs utilizing two different reprogramming constructs. All such iPSC-MSCs exhibited the characteristics of normal bone marrow-derived (BM) MSCs. In direct comparison to BM-MSCs our iPSC-MSCs exhibited a similar surface marker expression profile but shorter doubling times without reaching senescence within 20 passages. Considering functional capabilities, iPSC-MSCs provided supportive feeder layer for CD34+hematopoietic stem cells’ self-renewal and colony forming capacities. Furthermore, iPSC-MSCs gained immunomodulatory function to suppress CD4+cell proliferation, reduce proinflammatory cytokines in mixed lymphocyte reaction, and increase regulatory CD4+/CD69+/CD25+T-lymphocyte population. In conclusion, we generated fully functional MSCs from various iPSC lines irrespective of their starting cell source or reprogramming factor composition and we suggest that such iPSC-MSCs allow repetitive cell applications for advanced therapeutic approaches.

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