Expression Of Related Markers Research Articles

Isolation and Characterization of Human Dental Pulp Derived Stem Cells by Using Media Containing Low Human Serum Percentage as Clinical Grade Substitutes for Bovine Serum

Adult stem cells have been proposed as an alternative to embryonic stem cells to study multilineage differentiation in vitro and to use in therapy. Current culture media for isolation and expansion of adult stem cells require the use of large amounts of animal sera, but animal-derived culture reagents give rise to some questions due to the real possibility of infections and severe immune reactions. For these reasons a clinical grade substitute to animal sera is needed. We tested the isolation, proliferation, morphology, stemness related marker expression, and osteoblastic differentiation potential of Dental Pulp Stem Cells (DPSC) in a chemically defined medium containing a low percentage of human serum, 1.25%, in comparison to a medium containing 10% Fetal Bovine Serum (FBS). DPSCs cultured in presence of our isolation/proliferation medium added with low HS percentage were obtained without immune-selection methods and showed high uniformity in the expression of stem cell markers, proliferated at higher rate, and demonstrated comparable osteoblastic potential with respect to DPSCs cultured in 10% FBS. In this study we demonstrated that a chemically defined medium added with low HS percentage, derived from autologous and heterologous sources, could be a valid substitute to FBS-containing media and should be helpful for adult stem cells clinical application.

Abstract 1717: Prognostic impact of cancer stem cell-related markers in non-small cell lung cancer patients treated with induction chemoradiotherapy

Abstract Background: Several cancer stem cell (CSC) related markers have been confirmed to be expressed in non-small cell lung cancer (NSCLC). The aim of this study was to clarify the prognostic role of CSC related markers expression in locally advanced NSCLC with induction chemoradiotherap (CRT).Methods: Fifty patients with clinically N2 or N3 NSCLC underwent induction CRT with cisplatin and docetaxel concurrently with thoracic radiation followed by surgery. Expressions of CSC related markers (CD133, ABCG2, ALDH1, and Bmi-1) were examined by immunohistochemical stain from surgical resected specimen. The prognostic factors were investigated by multivariate analysis using Cox proportional hazard model.Results: Among 50 patients, 20 patients had no pathologically residual tumor cell in the surgical resected specimen, and CSC related markers expressions were evaluated in other 30 patients. With a median follow-up duration of 71 months, the 5-year overall survival rate in the patients of CD133 or ALDH1 -positive was significantly worse than that in the patients of CD133 and ALDH1 -negative (90.0% vs. 44.9%, respectively, P = 0.042). In multivariate analysis, CD133 and ALDH1 negativity (Hazard ratio [HR] 0.16, 95% confidential interval [CI]: 0.0086-0.98, P = 0.047) and c-N2-3 single node metastasis (not multi node metastasis) (HR 0.19, 95% CI: 0.028-0.91, P = 0.03) were significant independent prognostic factors for the prolongation of survival.Discussion: The expressions of CSC related markers after CRT are significantly related to poor prognosis in NSCLC. The development of therapeutic strategy including adjuvant therapy, is a key for further improvement of prognosis in patients who underwent trimodality therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1717. doi:1538-7445.AM2012-1717

Subcellular Distribution and Relative Expression of Fibrocyte Markers in the CD/1 Mouse Cochlea Assessed by Semiquantitative Immunogold Electron Microscopy

Spiral ligament fibrocytes function in cochlear homeostasis, maintaining the endocochlear potential by participating in potassium recycling, and fibrocyte degeneration contributes to hearing loss. Their superficial location makes them amenable to replacement by cellular transplantation. Fibrocyte cultures offer one source of transplantable cells, but determining what fibrocyte types they contain and what phenotype transplanted cells may adopt is problematic. Here, we use immunogold electron microscopy to assess the relative expression of markers in native fibrocytes of the CD/1 mouse spiral ligament. Caldesmon and aquaporin 1 are expressed more in type III fibrocytes than any other type. S-100 is strongly expressed in types I, II, and V fibrocytes, and α1Na,K-ATPase is expressed strongly only in types II and V. By combining caldesmon or aquaporin 1 with S-100 and α1Na,K-ATPase, a ratiometric analysis of immunogold density distinguishes all except type II and type V fibrocytes. Other putative markers (creatine kinase BB and connective tissue growth factor) did not provide additional useful analytical attributes. By labeling serial sections or by double or triple labeling with combinations of three antibodies, this technique could be used to distinguish all except type II and type V fibrocytes in culture or after cellular transplantation into the lateral wall.

Human hepatocellular carcinomas with “Stemness”-related marker expression: keratin 19 expression and a poor prognosis

There is a recently proposed subtype of hepatocellular carcinoma (HCC) that is histologically similar to usual HCC, but characterized by the expression of "stemness"-related markers. A large-scale study on two different cohorts of HCCs was performed to investigate the clinicopathologic features and epithelial-mesenchymal transition (EMT)-related protein expression status of this subtype of HCCs. The expression status of stemness-related (e.g., keratin 19 [K19], cluster of differentiation [CD]133, epithelial cell adhesion molecule [EpCAM], and c-kit) and EMT-related markers (e.g., snail, S100A4, urokinase plasminogen activator receptor [uPAR], ezrin, vimentin, E-cadherin, and matrix metalloproteinase [MMP]2) were examined using tissue microarrays from cohort 1 HCCs (n = 137). K19 protein expression in cohort 2 HCCs (n = 237) was correlated with the clinicopathologic parameters and messenger RNA (mRNA) levels of K19, uPAR, VIL2, Snail, Slug, and Twist. K19, EpCAM, c-kit, and CD133 positivity were observed in 18.2%, 35.0%, 34.3%, and 24.8%, respectively. K19 was most frequently expressed in combination with at least one other stemness-related marker (92.0%). K19-positive HCCs demonstrated more frequent major vessel invasion and increased tumor size, compared to K19-negative HCCs (P < 0.05). K19 was most significantly associated with EMT-related protein expression (e.g., vimentin, S100A4, uPAR, and ezrin) (P < 0.05) and a poor prognosis (overall survival: P = 0.018; disease-free survival: P = 0.007) in cohort 1. In cohort 2, HCCs with high K19 mRNA levels demonstrated higher mRNA levels of Snail, uPAR, and MMP2 (P < 0.05). K19-positive HCCs demonstrated more frequent microvascular invasion, fibrous stroma, and less tumor-capsule formation, compared to K19-negative HCCs (P < 0.05). K19 expression was a significant independent predictive factor of poor disease-free survival (P = 0.032). K19 was well correlated with clinicopathologic features of tumor aggressiveness, compared to other stemness-related proteins. K19-positive HCCs showed significantly increased EMT-related protein and mRNA expression, suggesting that they may acquire more invasive characteristics, compared to K19-negative HCCs through the up-regulation of EMT-associated genes.

Soleus muscle of Down syndrome (DS) mice exhibits elevated markers of oxidative and nitrosative stress

Individuals with DS have a reduced capacity for voluntary force generation compared to individuals with and without other forms of intellectual disability. Oxidative stress (OS) is thought to be a key factor in DS pathologies that may be the result of triplication and over‐expression of copper‐zinc superoxide dismutase (CuZnSod). Our aim was to assess markers of OS and nitrosative stress (NS) in soleus muscle of a well established mouse model of DS, the Ts65Dn mouse, which is segmental trisomic for CuZnSod. We hypothesized that soleus muscle of Ts65Dn mice would exhibit elevated markers of OS and NS compared to soleus from wild‐type (WT) controls. Eighteen (n=9 per group) 4–7 month old male Ts65Dn and WT mice were used in these experiments. Western blot analysis was performed to quantify the relative expression of markers of OS (4‐hydroxy‐nonenal (4‐HNE)) and NS (3‐nitrotryosine (3‐NT)). Levels of soleus muscle 4‐HNE and 3‐NT were 476% and 276% higher in Ts65Dn mice (p&lt;0.05), respectively. These findings suggest that soleus muscle from Ts65Dn mice exhibits elevated markers of OS and NS, which is consistent with the human phenotype of DS. Additionally, preliminary microarray analysis identified 43 up‐regulated and 66 down‐regulated genes in soleus of Ts65Dn compared to WT mice (p&lt;0.001, ± 1.5‐fold). We are currently categorizing these differentially expressed genes.Supported by Syracuse University SOE