Identification of Specific Cell-Surface Markers of Adipose-Derived Stem Cells from Subcutaneous and Visceral Fat Depots

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.

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Laminar and region‐specific cell surface markers in the entorhinal cortex and hippocampus

Hippocampus ◽

10.1002/hipo.1993.4500030722 ◽

1993 ◽

Vol 3 (S1) ◽

pp. 183-189 ◽

Cited By ~ 1

Author(s):

Peter L. Woodhams

Keyword(s):

Cell Surface ◽

Entorhinal Cortex ◽

Specific Cell ◽

Surface Markers ◽

Cell Surface Markers ◽

Specific Cell Surface

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Spherical self-organizing map as a helpful tool to identify category-specific cell surface markers

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2008.09.010 ◽

2008 ◽

Vol 376 (2) ◽

pp. 414-418 ◽

Cited By ~ 7

Author(s):

Tuoya ◽

Yuh Sugii ◽

Hitomi Satoh ◽

Dongwei Yu ◽

Yasaburo Matsuura ◽

...

Keyword(s):

Cell Surface ◽

Specific Cell ◽

Surface Markers ◽

Cell Surface Markers ◽

Self Organizing Map ◽

Helpful Tool ◽

Specific Cell Surface ◽

Self Organizing

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Abstract 16226: Single-cell RNA Sequencing Coupled With Optical Imaging for Targeted Real-time Visualization of the Cardiac Conduction System

Circulation ◽

10.1161/circ.142.suppl_3.16226 ◽

2020 ◽

Vol 142 (Suppl_3) ◽

Author(s):

William R Goodyer ◽

Benjamin Beyersdorf ◽

Nynke Van Den Berg ◽

Nazan Puluca ◽

Jan Buikema ◽

...

Keyword(s):

Cell Surface ◽

Optical Imaging ◽

Real Time ◽

Cardiac Conduction ◽

Specific Cell ◽

Surface Markers ◽

Cell Surface Markers ◽

Real Time Visualization ◽

Specific Cell Surface

Introduction: Optical imaging has the potential to revolutionize cardiothoracic surgery by allowing for the real-time visualization of structures often inadvertently damaged due to inadequate visibility. The cardiac conduction system (CCS) consists of specialized cells embedded within the heart that are essential for cardiac function yet indistinguishable from heart muscle tissue. Intraoperative CCS injury is a major complication in cardiac surgery, representing a significant source of morbidity and mortality. To date, there exists no intraoperative method to visualize the CCS. Hypothesis: We hypothesized that unique, CCS-specific cell surface markers could be used for the in vivo labelling of the CCS. Objectives: Use single-cell RNA sequencing (scRNAseq) to discover cell surface markers that may serve as the basis for generating optical imaging agents for real-time CCS visualization. Methods/Results: Gene expression analysis of a comprehensive scRNAseq dataset of the entire murine CCS revealed significant enrichment of a host of CCS-specific cell surface genes. A subset of genes were subsequently validated in the CCS of mice and/or human tissue. In total, 7 novel cell surface markers were confirmed to have unique expression patterns throughout or within distinct components of the CCS. Next, optical imaging agents were created consisting of a near-infrared (NIR) dye conjugated to antibodies directed against two distinct CCS-specific cell surface markers. Each optical imaging agent demonstrated high sensitivity and specificity in labeling the entire CCS in vivo following a single intravenous injection in mice. Specificity was confirmed within intact, whole hearts using both closed-field NIR imaging and whole mount immunolabeling with volume imaging (iDISCO+). Dosage, timecourse and biodistribution analyses were performed as well as safety validation by surface ECG. Conclusions: In summary, we coupled scRNAseq with optical imaging to create novel tools for the real-time visualization of a complex tissue substructure. We provide a proof-of-principle for broadening the scope of optical imaging but also address a significant unmet clinical need, laying the foundation for translational opportunities in cardiac intervention and imaging.

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Two Distinct Populations of Enriched Non Mobilized Peripheral Blood Mononuclear Cells (PBMNCs) with Different Functional Capacities

Blood ◽

10.1182/blood.v112.11.5402.5402 ◽

2008 ◽

Vol 112 (11) ◽

pp. 5402-5402

Author(s):

Nira Varda-Bloom ◽

Tatiana Kniazhansky ◽

Arnon Nagler ◽

Avraham J Treves

Keyword(s):

Stem Cells ◽

Cell Surface ◽

Peripheral Blood ◽

Ex Vivo ◽

Specific Cell ◽

Surface Markers ◽

Adherent Cells ◽

Cell Surface Markers ◽

Differentiation Potential ◽

Mobilized Peripheral Blood

Abstract Pluripotent stem cells are valuable sources for transplantation and tissue repair. The bone-marrow, umbilical cord blood and G-CSF mobilized peripheral blood are the main sources for adult stem cells. Non-mobilized peripheral blood contains mostly committed cells but recent studies suggest the presence of early progenitor and stem cells as well. Here we aim to develop a method to enrich and recover functional progenitor cell populations from non-mobilized peripheral blood. The ex-vivo enriched non-mobilized PBMNCs were tested by FACS analysis of specific cell surface markers, and by functional analysis of differentiation into different cells lineages. Non mobilized PBMNCs obtained from consenting healthy donors (n=18) cultured for 7 days in a defined cytokine cocktail media were analyzed by FACS using CD14, CD31, CD34, CD45, CD90, CD105, CD117 and CD133 and compare to non-manipulated PBMNCs. The enriched cells were also tested for their differentiation capacity into hematopoietic, endothelial and mesenchymal cells lineages compared to non-manipulated PBMNCs. We found that the enriched PBMNCs resulted in two distinct subpopulations, adherent and non adherent, that present different cell surface markers and have different differentiation capacities. Cell surface markers analysis showed that adherent cells possess high percentage of CD14 (28.1 ±6.13, 11.8 ± 3.9), CD90 (4.24±0.94, 1.53 ± 0.28), CD105 (42.19±8.42, 7.96 ± 4.8), CD117 (9.89±5.99, 1.75± 0.8) expression and reduction of CD45 (46.3±8.14, 62.74± 9.6), CD31 (5.9±1.9, 15.34 ± 4.9) and CD34 (0.18±0.03, 0.62 ±0.3) compare to non-manipulated PBMNCs, respectively. On the other hand, the non-adherent sub-population expresses more CD34 (1.2±0.02, 0.16±0.02), KDR (3.62±0.82, 0.49±0.2), CD105 (21.62±1.85, 7.96 ± 4.8) and CD45 (88.7±0.51, 62.74± 9.6) compared to non-manipulated PBMNCs, respectively. The adherent cells subpopulation showed higher differentiation potential into endothelial (116+23.1 EC colonies/106 cells), mesenchymal (400±53.9 CFU-F/106 cells) lineages compare to non-adherent cells (4.9±1.7 EC colonies/106 cells, 200±29.5 CFU-F/106 cells) and compare to non-manipulated PBMNCs (11.4±1.4 EC colonies/106 cells and 2.5±0.08 CFU-F/106 cells). The non-adherent subpopulations showed higher differentiation potential into hematopoietic colonies (445±75 CFU/106 cells) compared to the adherent sub-population (41.2±24.8 CFU/106 cells), and compared to the non-manipulated PBMNCs (373±39.7 CFU/106 cells) of total colony numbers/106 cells. All results (n=18) are presented as (mean ± SE). In summary, our ex-vivo enrichment methodology yields two different subpopulations with enriched hematological lineage in the non-adherent fraction and enriched endothelial and mesenchymal lineages in the adherent fraction. The ability to obtain enriched populations of endothelial, mesenchymal and hematopoietic progenitors from non mobilized peripheral blood cells may have an important clinical application.

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Neonatal stem cells exhibit specific characteristics in function, proliferation, and cellular signaling that distinguish them from their adult counterparts

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00031.2008 ◽

2008 ◽

Vol 294 (5) ◽

pp. R1491-R1497 ◽

Cited By ~ 14

Author(s):

Troy A. Markel ◽

Meijing Wang ◽

Paul R. Crisostomo ◽

Maiuxi C. Manukyan ◽

Jeffrey A. Poynter ◽

...

Keyword(s):

Stem Cells ◽

Cell Surface ◽

Surface Markers ◽

Cell Surface Markers ◽

Local Inflammatory Response ◽

Cell Counts ◽

Marrow Mesenchymal Stem Cells ◽

Growth Factor Production ◽

Erk Inhibition ◽

Different Levels

Stem cells may be a novel treatment modality for organ ischemia, possibly through beneficial paracrine mechanisms. Stem cells from older hosts have been shown to exhibit decreased function during stress. We therefore hypothesized that 1) neonatal bone marrow mesenchymal stem cells (nBMSCs) would produce different levels of IL-6, VEGF, and IGF-1 compared with adults (aBMSCs) when stimulated with TNF or LPS; 2) differences in cytokines would be due to distinct cellular characteristics, such as proliferation or pluripotent potential; and 3) differences in cytokines would be associated with differences in p38 MAPK and ERK signaling within nBMSCs. BMSCs were isolated from adult and neonatal mice. Cells were exposed to TNF or LPS with or without p38 or ERK inhibition. Growth factors were measured via ELISA, proliferation via daily cell counts, cell surface markers via flow cytometry, and pluripotent potential via alkaline phosphatase activity. nBMSCs produced lower levels of IL-6 and VEGF, but higher levels of IGF-1 under basal conditions, as well as after stimulation with TNF, but not LPS. Neonatal and adult BMSCs had similar pluripotent potentials and cell surface markers, but nBMSCs proliferated faster. Furthermore, p38 and ERK appeared to play a more substantial role in nBMSC cytokine and growth factor production. Neonatal stem cells may aid in decreasing the local inflammatory response during ischemia, and could possibly be expanded more rapidly than adult cells prior to therapeutic use.

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