scholarly journals Location and Clonal Analysis of Stem Cells and Their Differentiated Progeny in the Human Ocular Surface

1999 ◽  
Vol 145 (4) ◽  
pp. 769-782 ◽  
Author(s):  
Graziella Pellegrini ◽  
Osvaldo Golisano ◽  
Patrizia Paterna ◽  
Alessandro Lambiase ◽  
Stefano Bonini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Life History ◽  
Ocular Surface ◽  
Goblet Cells ◽  
Clonal Analysis ◽  
Proliferative Potential ◽  
Conjunctival Epithelium ◽  
Proliferative Capacity ◽  
Differentiation Potential ◽  
Goblet Cell Differentiation

We have analyzed the proliferative and differentiation potential of human ocular keratinocytes. Holoclones, meroclones, and paraclones, previously identified in skin, constitute also the proliferative compartment of the ocular epithelium. Ocular holoclones have the expected properties of stem cells, while transient amplifying cells have variable proliferative potential. Corneal stem cells are segregated in the limbus, while conjunctival stem cells are uniformly distributed in bulbar and forniceal conjunctiva. Conjunctival keratinocytes and goblet cells derive from a common bipotent progenitor. Goblet cells were found in cultures of transient amplifying cells, suggesting that commitment for goblet cell differentiation can occur late in the life of a single conjunctival clone. We found that conjunctival keratinocytes with high proliferative capacity give rise to goblet cells at least twice in their life and, more importantly, at rather precise times of their life history, namely at 45–50 cell doublings and at ∼15 cell doublings before senescence. Thus, the decision of conjunctival keratinocytes to differentiate into goblet cells appears to be dependent upon an intrinsic “cell doubling clock.” These data open new perspectives in the surgical treatment of severe defects of the anterior ocular surface with autologous cultured conjunctival epithelium.

The Influence of Oxygen on the Proliferative Capacity and Differentiation Potential of Lacrimal Gland–Derived Mesenchymal Stem Cells

2015 ◽  
Vol 56 (8) ◽  
pp. 4741 ◽  
Author(s):  
Mathias Roth ◽  
Kristina Spaniol ◽  
Claus Kordes ◽  
Silke Schwarz ◽  
Sonja Mertsch ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Lacrimal Gland ◽  
Proliferative Capacity ◽  
Differentiation Potential

Poor Potential of Proliferation and Differentiation in Bone Marrow Mesenchymal Stem Cells Derived From Children with Severe Aplastic Anemia.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 5081-5081
Author(s):  
Ching-Tien Peng
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Aplastic Anemia ◽  
Severe Aplastic Anemia ◽  
Population Doubling ◽  
Bone Marrow Mscs ◽  
Proliferative Capacity ◽  
Differentiation Potential ◽  
Proliferation And Differentiation

Abstract 5081 Introduction Idiopathic severe aplastic anemia (SAA), characterized by failure of hematopoiesis, is rare and potentially life-threatening to children. However, the pathogenesis has not been completely understood, and insufficiency in the hematopoietic microenvironment can be an important factor. Mesenchymal stem cells (MSCs) play an important role in maintaining bone marrow microenvironment. Therefore, we aimed at the intrinsic defects of bone marrow MSCs derived from SAA children. Materials and Methods Bone marrow MSCs were obtained from 5 SAA children and 5 controls. The morphology, immunophenotyping, proliferative capacity and differentiation potential of MSCs from SAA children were determined and compared with those of MSCs from controls. Results In vitro, MSCs of SAA and control group shared a similar spindle-shaped morphology. Both revealed a consistent immunophenotypic profile which was negative for CD45, CD14 and CD34, and positive for CD105, CD73, and CD44. However, SAA MSCs had slower expansion rate and smaller cumulative population doubling from passage 4 to 6 (1.83± 1.21 vs 3.36± 0.87; p = 0.046), indicating lower proliferative capacity. Besides, only 3 of 5 cultures of SAA group retained the ability to continue expansion till 80%-90% confluent cell layer beyond passage 6, suggesting earlier senescence of SAA MSCs. After osteogenic induction, SAA MSCs showed lower alkaline phosphatase activity (1.46± 0.04 vs 2.27± 0.32; p = 0.013), less intense von Kossa staining and lower gene expression of core binding factora1 (0.0015± 0.0005 vs 0.0056± 0.0017; p = 0.013). Following adipogenic induction, SAA MSCs showed less intense Oil red O staining (0.86± 0.22 vs 1.73± 0.42; p = 0.013) and lower lipoproteinlipase expression (0.0105± 0.0074 vs 0.0527± 0.0254; p = 0.013).The results of real time-PCR analysis for the assessment of lineage-specific genes were consistent with the findings of histochemical stains, and both indicated that SAA MSCs had poor osteogenic and adipogenic potential. Conclusions In this study, we demonstrated that bone marrow MSCs from children with SAA had poor potential of proliferation and differentiation. These alterations in MSCs may contribute to the failure of hematopoiesis, and lead to the development of the disease. Further studies are needed to elucidate the relationship between MSCs and SAA. Disclosures No relevant conflicts of interest to declare.

scholarly journals Treprostinil indirectly regulates endothelial colony forming cell angiogenic properties by increasing VEGF-A produced by mesenchymal stem cells

2015 ◽  
Vol 114 (10) ◽  
pp. 735-747 ◽  
Author(s):  
Marilyne Levy ◽  
Lan Huang ◽  
Elisa Rossi ◽  
Adeline Blandinières ◽  
Dominique Israel-Biet ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cord Blood ◽  
Proliferative Potential ◽  
Differentiation Potential ◽  
Endothelial Colony Forming Cells ◽  
Pulmonary Vasodilators ◽  
High Level

SummaryPulmonary vasodilators and prostacyclin therapy in particular, have markedly improved the outcome of patients with pulmonary hypertension (PH). Endothelial dysfunction is a key feature of PH, and we previously reported that treprostinil therapy increases number and proliferative potential of endothelial colony forming cells (ECFC) isolated from PH patients’ blood. In the present study, the objective was to determine how treprostinil contributes to the proangiogenic functions of ECFC. We examined the effect of treprostinil on ECFC obtained from cord blood in terms of colony numbers, proliferative and clonogenic properties in vitro, as well as in vivo vasculogenic properties. Surprisingly, treprostinil inhibited viability of cultured ECFC but did not modify their clonogenic properties or the endothelial differentiation potential from cord blood stem cells. Treprostinil treatment significantly increased the vessel-forming ability of ECFC combined with mesenchymal stem cells (MSC) in Matrigel implanted in nude mice. In vitro, ECFC proliferation was stimulated by conditioned media from treprostinil-pretreated MSC, and this effect was inhibited either by the use of VEGF-A blocking antibodies or siRNA VEGF-A in MSC. Silencing VEGF-A gene in MSC also blocked the pro-angiogenic effect of treprostinil in vivo. In conclusion, increased VEGF-A produced by MSC can account for the increased vessel formation observed during treprostinil treatment. The clinical relevance of these data was confirmed by the high level of VEGF-A detected in plasma from patients with paediatric PH who had been treated with treprostinil. Moreover, our results suggest that VEGF-A level in patients could be a surrogate biomarker of treprostinil efficacy.

Clonal analysis of the differentiation potential of human adipose-derived adult stem cells

2005 ◽  
Vol 206 (1) ◽  
pp. 229-237 ◽  
Author(s):  
Farshid Guilak ◽  
Kristen E. Lott ◽  
Hani A. Awad ◽  
Qiongfang Cao ◽  
Kevin C. Hicok ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adult Stem Cells ◽  
Clonal Analysis ◽  
Differentiation Potential

scholarly journals Urine-Derived Stem Cells: Differentiation Potential into Smooth-Muscle Cells and Urothelial Cell

2019 ◽  
Vol 74 (3) ◽  
pp. 176-184
Author(s):  
Igor A. Vasyutin ◽  
Aleksey V. Lyundup ◽  
Sergey L. Kuznetsov
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Smooth Muscle ◽  
Urinary Tract ◽  
Urinary Bladder ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Proliferative Potential ◽  
Differentiation Potential ◽  
Invasive Approach

Background: Tissue engineering of low urinary tract organs requires biopsy of urinary bladder material. The current study describes non-invasive approach of obtaining autologous stem cells from urine of healthy adults. These cells were studied for potential to differentiate into epithelial cells and smooth muscle cells of the urinary bladder. Aims: To describe properties of urine-derived stem cells (USCs) and investigate their differentiation potential for tissue engineering of low urinary tract organs. Materials and Methods: USCs were isolated from urine of healthy volunteers with centrifugation and seeded in media to 24-well plates. Expression of stem cells markers (CD73, CD90, CD105, CD34, CD45, CD29, CD44, CD54, SSEA4) by USCs was assessed with flow cytometry. Expression of specific markers of smooth muscle cells and urothelial cells was assessed with fluorescence microscopy with following computational image analysis. Results: Median number of USCs per 100 ml urine was 6. Doubling time for USC was 1.440.528 days (n=4) and there were 26.34.79 population doublings for USC cultures (n=4). Median expression of markers of postnatal stem cells was CD73 ― 79.8%, CD90 ― 56.6%, CD105 ― 40.7%, CD34 1.0%, CD45 2.0%, CD29 99.0%, CD44 99.0%, CD54 ― 97.7% and SSEA4 99.0%. Treatment of cells with high concentration of EGF in media with low concentration of FBS for 10 days increased cytokeratin (CK) expression to 24.9% for CK AE1/AE3 and to 7.6% for CK 7. Treatment of USCs with media inducing smooth muscle differentiation for 10 days increased expression of -smooth muscle actin to 79.6% and expression of calponin to 97.6%. Conclusions: USCs are cells that can be found in urine in small quantities. They have high proliferative potential and express markers of postnatal stem cells. Under effect of PDGF-BB and TGF- 1 they differentiate into smooth muscle cells.

scholarly journals Adult neural stem cells have latent inflammatory potential that is kept suppressed by Tcf4 to facilitate adult neurogenesis

2021 ◽  
Vol 7 (21) ◽  
pp. eabf5606
Author(s):  
Mohammad Shariq ◽  
Vinaya Sahasrabuddhe ◽  
Sreevatsan Krishna ◽  
Swathi Radha ◽  
Nruthyathi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Dentate Gyrus ◽  
Adult Neurogenesis ◽  
Neural Progenitors ◽  
The Other ◽  
Proliferative Capacity ◽  
Differentiation Potential ◽  
Neurogenic Niche ◽  
Transcription Factor 4

Inflammation is known to adversely affect adult neurogenesis, wherein the source of inflammation is largely thought to be extraneous to the neurogenic niche. Here, we demonstrate that the adult hippocampal neural progenitors harbor an inflammatory potential that is proactively suppressed by transcription factor 4 (Tcf4). Deletion of Tcf4 in hippocampal nestin-expressing progenitors causes loss of proliferative capacity and acquisition of myeloid inflammatory properties. This transformation abolishes their differentiation potential and causes production of detrimental factors that adversely affect niche cells, causing inflammation in the dentate gyrus. Thus, on one hand, Tcf4 deletion causes abrogation of proliferative progenitors leading to reduction of adult neurogenesis, while on the other, their accompanying inflammatory transformation inflicts inflammation in the niche. Taken together, we provide the first evidence for a latent inflammatory potential of adult hippocampal neural progenitors and identify Tcf4 as a critical regulator that facilitates adult neurogenesis via proactive suppression of this detrimental potential.

scholarly journals Conjunctival Goblet Cell Responses to TLR5 Engagement Promote Activation of Local Antigen-Presenting Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Abiramy Logeswaran ◽  
Laura Contreras-Ruiz ◽  
Sharmila Masli
Keyword(s):  
Dendritic Cells ◽  
Goblet Cell ◽  
Ocular Surface ◽  
Primary Cultures ◽  
Goblet Cells ◽  
Microbial Colonization ◽  
Mucosal Inflammation ◽  
Conjunctival Epithelium ◽  
Cell Responses ◽  
Mucosal Homeostasis

Conjunctival epithelium forms a barrier between the ocular surface microbial flora and the ocular mucosa. In addition to secreting gel-forming mucins, goblet cells, located in the conjunctival epithelium, help maintain local immune homeostasis by secreting active TGFβ2 and promoting tolerogenic phenotype of dendritic cells in the vicinity. Although dendritic cell subsets, characteristic of mucosal tissues, are found in the conjunctiva, previous studies provided limited information about their location within the tissue. In this study, we examine immunostained conjunctiva explants to determine the location of CD11c-positive dendritic cells in the context of MUC5AC-positive goblet cells. Considering that conjunctival goblet cells are responsive to signaling induced by pathogen recognition receptors, we also assess if their responses to microbial product, flagellin, can contribute to the disruption of ocular mucosal homeostasis that promotes activation of dendritic cells and results in chronic ocular surface inflammation. We find that dendritic cells in the conjunctiva with an increased microbial colonization are located adjacent to goblet cells. While their cell bodies in the stromal layer are immediately below the epithelial layer, several extensions of dendritic cells are projected across the epithelium towards the ocular surface. Such trans-epithelial dendrites are not detectable in healthy ocular mucosa. In response to topically applied flagellin, increased proportion of CD11c-positive cells in the conjunctiva strongly express MHC class II relative to the untreated conjunctiva. This change is accompanied by reduced immunoreactivity to TGFβ-activating Thrombospondin-1 in the conjunctival epithelium. These findings are supported by in vitro observations in primary cultures of goblet cells that respond to the TLR5 stimulation with an increased expression of IL-6 and reduced level of active TGFβ. The observed changes in the conjunctiva after flagellin application correspond with the development of clinical signs of chronic ocular mucosal inflammation including corneal epitheliopathy. Collectively, these findings demonstrate the ability of ocular mucosal dendritic cells to extend trans-epithelial dendrites in response to increased microbial colonization at the ocular surface. Moreover, this study provides key insight into how goblet cell responses to microbial stimuli may contribute to the disruption of ocular mucosal homeostasis and chronic ocular mucosal inflammation.

scholarly journals Molecular Aspects of Adipose-Derived Stromal Cell Senescence in a Long-Term Culture: A Potential Role of Inflammatory Pathways

2020 ◽  
Vol 29 ◽  
pp. 096368972091734 ◽  
Author(s):  
Marta Pokrywczynska ◽  
Małgorzata Maj ◽  
Tomasz Kloskowski ◽  
Monika Buhl ◽  
Daria Balcerczyk ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Doubling Time ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Proliferative Potential ◽  
Differentiation Potential ◽  
Long Term Culture ◽  
Stromal Stem Cells

Long-term culture of mesenchymal stromal/stem cells in vitro leads to their senescence. It is very important to define the maximal passage to which the mesenchymal stromal/stem cells maintain their regenerative properties and can be used for cellular therapies and construction of neo-organs for clinical application. Adipose-derived stromal/stem cells were isolated from porcine adipose tissue. Immunophenotype, population doubling time, viability using bromodeoxyuridine assay, MTT assay, clonogencity, β-galactosidase activity, specific senescence-associated gene expression, apoptosis, and cell cycle of adipose-derived mesenchymal stromal/stem cells (AD-MSCs) were analyzed. All analyses were performed through 12 passages (P). Decreasing viability and proliferative potential of AD-MSCs with subsequent passages together with prolonged population doubling time were observed. Expression of β-galactosidase gradually increased after P6. Differentiation potential of AD-MSCs into adipogenic, chondrogenic, and osteogenic lineages decreased at the end of culture (P10). No changes in the cell cycle, the number of apoptotic cells and expression of specific AD-MSC markers during the long-term culture were revealed. Molecular analysis showed increased expression of genes involved in activation of inflammatory response. AD-MSCs can be cultured for in vivo applications without loss of their properties up to P6.

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