scholarly journals Phenotype Characterization of Human Adipose-Derived Stem Cell by Flow Cytometry

2021 ◽  
Author(s):  
Celeste Limoli ◽  
Paolo Giuseppe Limoli ◽  
Marcella Nebbioso
Keyword(s):  
Adipose Tissue ◽  
Cell Surface ◽  
Stromal Vascular Fraction ◽  
Surface Markers ◽  
Adipose Derived Stem Cells ◽  
Cell Surface Markers ◽  
Cell Surface Antigens ◽  
Adipose Derived Stem Cell

Background: Developing an efficient and standardized method to isolate and characterize adipose-derived stem cells (ASCs) from the stromal vascular fraction (SVF) of the adipose tissue for clinical application represents one of the major challenges in cell therapy and tissue engineering. Methods: In this study, we proposed an innovative, non-enzymatic protocol to collect clinically useful ASCs within freshly isolated SVF from adipose tissue by centrifugation of the infranatant portion of lipoaspirate and to determine the characteristic cytofluorimetric pattern, prior to in vitro culture. Results: The SVF yielded a mean of 73,32 \pm\ 10,89% cell viability evaluated with CALCEINA-FITC, i.e. cell-permeant dye. The ASCs were positive for PC7-labeled mAb anti-CD34 and negative for both PE-labeled mAb anti-CD31 and APC-labeled mAb anti-CD45. The frequency of ASCs estimated according to the panel of cell surface markers used was 51,06%\ \pm 5,26% versus the unstained ASCs subpopulation that was 0,74%\pm0,84% (P<0.0001). The ASCs events/\muL were 1602,13/\muL \pm 731,87/\muL. Conclusion: Our findings suggested that ASCs found in freshly isolated adipose SVF obtained by centrifugation of lipoaspirate can be immunophenotypically identified with a basic panel of cell surface markers. These findings aimed to provide standardization and contribute to reducing the inconsistency on reported cell surface antigens of ASC derived from the existing literature.

scholarly journals In vitro Comparison of Adipogenic Differentiation in Human Adipose-Derived Stem Cells Cultured with Collagen Gel and Platelet-Rich Fibrin

2021 ◽  
Vol 54 (03) ◽  
pp. 278-283
Author(s):  
Pallavi Priyadarshini ◽  
Soumi Samuel ◽  
Basan Gowda Kurkalli ◽  
Chethan Kumar ◽  
Basavarajappa Mohana Kumar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Soft Tissue ◽  
Cell Surface ◽  
Adipogenic Differentiation ◽  
Collagen Gel ◽  
Surface Markers ◽  
Adipose Derived Stem Cells ◽  
Cell Surface Markers ◽  
Platelet Rich Fibrin

Abstract Background: Adipose-derived stem cells (ADSCs) are the most preferred cell type, based on their phenotypic characteristics, plasticity, and favorable immunological properties for applications in soft-tissue augmentation. Hence, the present in vitro study was aimed to evaluate the adipogenic differentiation potential of human ADSCs upon culturing individually with collagen gel and platelet-rich fibrin (PRF). Materials and methods: The collected lipoaspirate was used for establishing ADSCs using enzymatic digestion method. Then, the cells were analyzed for their morphology, viability, proliferation rate, population doubling time (PDT), colony-forming ability, cell surface markers expression, and osteogenic differentiation as biological properties. Further, ADSCs were evaluated for their adipogenicity using induction media alone, and by culturing with collagen gel and PRF individually for prospective tissue augmentation. Results: ADSCs were successfully established in vitro and exhibited a fibroblast-like morphology throughout the culture period. Cells had higher viability, proliferation potential and showed their ability to form colonies. The positive expression of cell surface markers and osteogenic ability confirmed the potency of ADSCs. The ADSCs cultured on collagen gel and PRF, individually, showed higher number of differentiated adipocytes than ADSCs grown with adipogenic induction medium alone. Conclusion: The extent of lipid accumulation by ADSCs was slightly higher when cultured on collagen gel than on PRF. Additional experiments are required to confirm better suitability of scaffold materials for soft-tissue regeneration.

Isolation and characterization of human CD34−Lin− and CD34+Lin− hematopoietic stem cells using cell surface markers AC133 and CD7

Blood ◽  
2000 ◽  
Vol 95 (9) ◽  
pp. 2813-2820 ◽  
Author(s):  
Lisa Gallacher ◽  
Barbara Murdoch ◽  
Dongmei M. Wu ◽  
Francis N. Karanu ◽  
Mike Keeney ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Hematopoietic Stem ◽  
Human Cd34 ◽  
Cd34 Cells

Recent evidence indicates that human hematopoietic stem cell properties can be found among cells lacking CD34 and lineage commitment markers (CD34−Lin−). A major barrier in the further characterization of human CD34− stem cells is the inability to detect this population using in vitro assays because these cells only demonstrate hematopoietic activity in vivo. Using cell surface markers AC133 and CD7, subfractions were isolated within CD34−CD38−Lin− and CD34+CD38−Lin− cells derived from human cord blood. Although the majority of CD34−CD38−Lin− cells lack AC133 and express CD7, an extremely rare population of AC133+CD7− cells was identified at a frequency of 0.2%. Surprisingly, these AC133+CD7− cells were highly enriched for progenitor activity at a frequency equivalent to purified fractions of CD34+ stem cells, and they were the only subset among the CD34−CD38−Lin− population capable of giving rise to CD34+ cells in defined liquid cultures. Human cells were detected in the bone marrow of non-obese/severe combined immunodeficiency (NOD/SCID) mice 8 weeks after transplantation of ex vivo–cultured AC133+CD7− cells isolated from the CD34−CD38−Lin− population, whereas 400-fold greater numbers of the AC133−CD7− subset had no engraftment ability. These studies provide novel insights into the hierarchical relationship of the human stem cell compartment by identifying a rare population of primitive human CD34− cells that are detectable after transplantation in vivo, enriched for in vitro clonogenic capacity, and capable of differentiation into CD34+ cells.

scholarly journals Identification of cell surface markers and establishment of monolayer differentiation to retinal pigment epithelial cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Alvaro Plaza Reyes ◽  
Sandra Petrus-Reurer ◽  
Sara Padrell Sánchez ◽  
Pankaj Kumar ◽  
Iyadh Douagi ◽  
...  
Keyword(s):  
Cell Surface ◽  
Retinal Pigment Epithelial ◽  
Retinal Pigment ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
In Vitro Differentiation ◽  
Rpe Cells ◽  
Pigment Epithelial ◽  
Differentiation Efficiency

AbstractIn vitro differentiation of human pluripotent stem cells into functional retinal pigment epithelial (RPE) cells provides a potentially unlimited source for cell based reparative therapy of age-related macular degeneration. Although the inherent pigmentation of the RPE cells have been useful to grossly evaluate differentiation efficiency and allowed manual isolation of pigmented structures, accurate quantification and automated isolation has been challenging. To address this issue, here we perform a comprehensive antibody screening and identify cell surface markers for RPE cells. We show that these markers can be used to isolate RPE cells during in vitro differentiation and to track, quantify and improve differentiation efficiency. Finally, these surface markers aided to develop a robust, direct and scalable monolayer differentiation protocol on human recombinant laminin-111 and −521 without the need for manual isolation.

Expression of cell surface markers during self-renewal and differentiation of human adipose-derived stem cells

2013 ◽  
Vol 430 (3) ◽  
pp. 871-875 ◽  
Author(s):  
Tala Mohsen-Kanson ◽  
Anne-Laure Hafner ◽  
Brigitte Wdziekonski ◽  
Phi Villageois ◽  
Bérengère Chignon-Sicard ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Surface ◽  
Surface Markers ◽  
Adipose Derived Stem Cells ◽  
Cell Surface Markers ◽  
Self Renewal

Prognostic Value of Immunophenotype Analysis in Patients with Acute Promyelocytic Leukemia (APL) Treated with All-Transretinoic Acid and Anthracyclin Monochemotherapy.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2345-2345
Author(s):  
Pau Montesinos ◽  
Concepcion Rivas ◽  
Consuelo Rayon ◽  
Edo Vellenga ◽  
Javier de la Serna ◽  
...  
Keyword(s):  
Cell Surface ◽  
Surface Antigens ◽  
Male Gender ◽  
Prognostic Impact ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Cell Surface Antigens ◽  
Hla Dr ◽  
And Gender ◽  
The Impact

Abstract Introduction: The prognostic significance of the expression pattern of certain cell surface markers in APL is controversial. Objectives: Analyse the impact of the expression of certain cell surface markers on complete remission rate (CR), overall survival (OS) and relapse free survival (RFS) in patients with APL included in multicenter trials PETHEMA LPA96 y LPA99. Material and methods: Between 1996 and 2005, 734 patients were included in these 2 consecutive trials. Induction therapy consisted of ATRA and idarubicin, followed by three consolidation courses of anthracycline monochemotherapy with or without ATRA and followed by maintenance. Bone marrow immunophenotype analysis was performed at local or reference laboratories. Positivity was defined as more than 20% blasts expressing a specific antigen for the following antigens: CD34 (527 patients), CD33 (521), CD15 (520), CD13 (513), HLA-DR (495), CD2 (443), CD19 (433), CD7 (403), CD117 (395), CD56 (392), y CD11b (335). We performed univariate analysis to establish the impact of antigen positivity on CR rate, OS and RFS. Significant values were included in the multivariate analysis. Results: A total of 664 patients (90%) achieved CR. The following variables were associated with decreased CR rate: WBC > 10x109/L, serum level creatinine > 1.4 mg/dl, age > 60 years, ECOG > 1, M3v and male gender. None of the cell surface antigens were significantly associated with CR rate. WBC, creatinine, age and gender were found to be independent prognostic factors for CR. Median follow up was 55 months. OS at 8 years was inferior in those patients with WBC > 10x109/L (67% vs 85%, p < 0.01), M3v (70% vs 83%, p < 0.01), age > 60 (56% vs 86%, p < 0.01), male gender (78% vs 83%, p=0.03), LPA96 trial (74% vs 84%, p=0.01) and CD2+ (76% vs 84%, p=0.04). Age, WBC and gender were independent factors for OS. RFS was inferior in those patients with WBC > 10x109/L (69% vs 93%, p < 0.01), high vs intermediate vs low risk (69% vs 91% vs 95%, p < 0.01), M3v (76% vs 88%, p < 0.01), BCR2 vs BCR3 vs BCR1 transcript (71% vs 81% vs 89%, p < 0.01), male gender (83% vs 90%, p=0.03), LPA96 trial (82% vs 87%, p=0.02) and CD2+ (75% vs 91%, p < 0.01). The risk of relapse category was the only independent factor for RFS. CD2+ APL (115/443 patients) was significantly associated with WBC > 10x109/L, M3v, BCR3, CD34+, CD56+, CD7+, and HLA-DR negative. Conclusion: Of all the cell surface antigens analysed, only expression of CD2 was associated with an lower OS and RFS, due to its association with WBC > 10x109/L. In patients taking part in PETHEMA trials, immunophenotype analysis at presentation does not give additional prognostic impact from the previously established risk factors.

MODULATION OF CELL SURFACE MARKERS ON NK-LIKE T LYMPHOCYTES BY USING IL-2, IL-7 OR IL-12 IN VITRO STIMULATION

Cytokine ◽  
2000 ◽  
Vol 12 (9) ◽  
pp. 1385-1390 ◽  
Author(s):  
B Zoll ◽  
P Lefterova ◽  
O Ebert ◽  
D Huhn ◽  
A von Ruecker ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cell Surface ◽  
Surface Markers ◽  
Cell Surface Markers

scholarly journals The role of cell surface markers and enamel matrix derivatives on human periodontal ligament mesenchymal progenitor responses in vitro

Biomaterials ◽  
2011 ◽  
Vol 32 (30) ◽  
pp. 7375-7388 ◽  
Author(s):  
Philippe Kémoun ◽  
Stan Gronthos ◽  
Malcolm L. Snead ◽  
Jacqueline Rue ◽  
Bruno Courtois ◽  
...  
Keyword(s):  
Cell Surface ◽  
Periodontal Ligament ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Enamel Matrix ◽  
Enamel Matrix Derivatives ◽  
Matrix Derivatives

scholarly journals Flow Cytometric Characterization of Cell Surface Markers to Differentiate Between Fibroblasts and Mesenchymal Stem Cells of Different Origin

2021 ◽  
Author(s):  
Suzanne Sober ◽  
Homa Darmani ◽  
Dana Alhattab ◽  
Abdalla Awidi
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Cell Surface ◽  
Placental Tissue ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Wharton's Jelly ◽  
Different Origins

IntroductionIdentification and purification of mesenchymal stem cells (MSCs) expanded in culture for therapeutic use is crucial for improved yield and optimal results. Fibroblasts are the most common cell type in connective tissue and are commonly found as contaminants of MSC cultures, affecting cell yield and potentially causing tumor formation after cell transplantation. In the current study, we wished to identify cell surface markers that can differentiate MSCs of different origins from fibroblasts.Material and methodsMSCs were isolated from bone marrow, adipose tissue, Wharton’s jelly and placental tissue and fibroblasts were isolated from foreskin (as a negative control) in order to examine the differences in the expression of a panel of 14 different cell surface markers using multiplex flow cytometry.ResultsOur results indicate that the following markers could be useful in differentiating between fibroblasts and MSCs derived from: adipose tissue - CD79a, CD105, CD106, CD146, and CD271; Wharton’s jelly - CD14, CD56 and CD105; bone marrow - CD105, CD106, and CD146; placental tissue - CD14, CD105, and CD146. Furthermore, we found that, contradictory to previous studies, CD26 is not fibroblast-specific.ConclusionsIn conclusion, the results of our study indicate that cell surface markers may prove to be a useful tool in the discrimination between MSCs of different origins and fibroblasts and thus may be used to authenticate the identity of the isolated cells.

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