Growth Properties Of Cells Research Articles

Design and synthesis of improved active-site SHP2 inhibitors with anti-breast cancer cell effects

The Src homology containing phosphotyrosyl phosphatase 2 (SHP2) is a bona fide oncogene particularly in cancers driven by overexpression of receptor tyrosine kinases (RTKs). As such, there is a growing interest to target SHP2 in cancer. Based on these premises, several active site (type I) and allosteric site (type II) inhibitors have been developed, but no SHP2 targeting therapies have reached the clinic yet. In an effort to fill these gaps, we embarked on producing optimized versions of our parent active-site SHP2 inhibitor CNBDA. The objectives were to produce derivatives with increased inhibitory potential and improved selectivity. Accordingly, we designed derivatives around the CNBDA scaffold and predicted their binding property by in silico molecular modeling. Based on comparative differences in free energy of binding to the SHP2 versus the SHP1 active sites, ten were selected, chemically synthesized, and evaluated by NMR and mass spectroscopy for structural integrity. Among the ten derivatives, BPDA2 was found to be the most potent and highly selective compound, inhibiting the SHP2 enzyme activity with an IC50 of 92 nM when DiFMUP was used as a substrate and with an IC50 of 47 nM when pNPP was used as a substrate. Furthermore, enzyme kinetic analyses showed that BPDA2 is a competitive SHP2 inhibitor. Selectivity comparisons in a PTPase assay using DiFMUP as a substrate demonstrated that BPDA2 is more selective to SHP2 than to SHP1 and PTP1B by more than 369-fold and 442-fold, respectively. Evaluation with a cellular thermal shift assay (CETSA) confirmed that BPDA2 binds to wild-type SHP2 in a cellular context, and stabilizes it in solution. Treatment of cells with DBDA2 downregulates mitogenic and cell survival signaling and RTK expression in a concentration dependent manner. Furthermore, treatment of cells with BPDA2 suppresses anchorage independent growth and cancer stem cell properties of breast cancer cells. Overall, data described in this report show that BPDA2 is a more potent derivative of CNBDA with a highly improved selectivity for SHP2.

Graphene Incorporated Electrospun Nanofiber for Electrochemical Sensing and Biomedical Applications: A Critical Review.

The extraordinary material graphene arrived in the fields of engineering and science to instigate a material revolution in 2004. Graphene has promptly risen as the super star due to its outstanding properties. Graphene is an allotrope of carbon and is made up of sp2-bonded carbon atoms placed in a two-dimensional honeycomb lattice. Graphite consists of stacked layers of graphene. Due to the distinctive structural features as well as excellent physico-chemical and electrical conductivity, graphene allows remarkable improvement in the performance of electrospun nanofibers (NFs), which results in the enhancement of promising applications in NF-based sensor and biomedical technologies. Electrospinning is an easy, economical, and versatile technology depending on electrostatic repulsion between the surface charges to generate fibers from the extensive list of polymeric and ceramic materials with diameters down to a few nanometers. NFs have emerged as important and attractive platform with outstanding properties for biosensing and biomedical applications, because of their excellent functional features, that include high porosity, high surface area to volume ratio, high catalytic and charge transfer, much better electrical conductivity, controllable nanofiber mat configuration, biocompatibility, and bioresorbability. The inclusion of graphene nanomaterials (GNMs) into NFs is highly desirable. Pre-processing techniques and post-processing techniques to incorporate GNMs into electrospun polymer NFs are precisely discussed. The accomplishment and the utilization of NFs containing GNMs in the electrochemical biosensing pathway for the detection of a broad range biological analytes are discussed. Graphene oxide (GO) has great importance and potential in the biomedical field and can imitate the composition of the extracellular matrix. The oxygen-rich GO is hydrophilic in nature and easily disperses in water, and assists in cell growth, drug delivery, and antimicrobial properties of electrospun nanofiber matrices. NFs containing GO for tissue engineering, drug and gene delivery, wound healing applications, and medical equipment are discussed. NFs containing GO have importance in biomedical applications, which include engineered cardiac patches, instrument coatings, and triboelectric nanogenerators (TENGs) for motion sensing applications. This review deals with graphene-based nanomaterials (GNMs) such as GO incorporated electrospun polymeric NFs for biosensing and biomedical applications, that can bridge the gap between the laboratory facility and industry.

Development and characterisation of 3D collagen-gelatin based scaffolds for breast cancer research

While 2D culture presents a useful tool for cancer research, it fails to replicate the tumor microenvironment as it lacks proper three-dimensional cell-cell/cell-matrix interactions, often resulting in exaggerated responses to therapeutic agents. 3D models that aim to overcome the issues associated with 2D culture research offer a new frontier for cancer research with cell growth, morphology and genetic properties that more closely match in vivo cancers. Herein, we aim to develop a collagen-based scaffold that supports the attachment and proliferation of breast cancer (BC) cells as a 3D culture model. Scaffolds were produced on a repeatable basis using a freeze-drying procedure. The constructs were highly porous (>99%) with homogenous pore sizes (150–300 μm) and an interconnected structure. The application of 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC) crosslinking resulted in scaffolds with elastic moduli in the range of 1–2 kPa, mimicking cancerous breast tissue stiffness. Furthermore, the incorporation of gelatin into the scaffolds enabled the porosity, pore size and mechanical properties to be tailored, resulting in scaffolds with stiffness values that accurately replicate the stiffness of human BC extracellular matrix (ECM) (1.3–1.7 kPa). Scaffolds displayed high in vitro stability with 90% of mass remaining after 14 days of culture. The scaffolds were shown to be highly biocompatible, and capable of supporting the attachment, infiltration and proliferation of MCF7 breast cancer (BC) cells over +14 days. These results confirm the suitability of these scaffolds as culture models for BC cells. These collagen-based scaffolds offer significant potential for the exploration of aspects of BC, such as gene expression profiles and patterns, and for the assessment of the efficacy of therapeutic agents in treating BC.

Development and characterization of Spodoptera mauritia ovarian primary cell culture and evaluation of fenoxycarb toxicity.

The rice swarming caterpillar, Spodoptera mauritia (Biosduval, 1833) (Lepidoptera: Noctuidae), has been considered a severe pest and has caused the extreme loss of rice paddies in the nursery stages. The massive reproductive efficiency of pests results from balanced endocrine functioning. Insect treatments with exogenous either juvenile hormones (JH) or juvenile hormone analogues (JHAs) during low endogenous JH titers disrupt metamorphosis andovarian development. Hence, it was thought worthwhile to develop a primary ovarian cell culture of Spodoptera mauritia to study the biological efficiency of JHA, fenoxycarb, at the cellular level. The study envisioned the cell characteristics and growth properties and found that most cells were spherical. Spindle-shaped cells were also present during the initial stage of active cell division. The majority of the cells grew attached to the bottom of the culture plates, and a few grew in suspension. The cell doubling time was 144 ± 14h. The growth curve exhibited a sigmoid pattern of cell proliferation at first, followed by a declining phase. These properties indicate that the primary ovarian culture is a suitable candidate for developing a novel cell line of S. mauritia. The preliminary study investigated the cytotoxicity of the insect growth regulator fenoxycarb in ovarian, primary cell culture. Incubated cells showed noticeable morphological changes, including cell shrinkage and apoptotic body formation; inhibition of cellular proliferation; and apoptosis-inducing effects in ovarian, primary cell culture of S. mauritia. Evidently, from these results, it is clear that juvenile hormone mimics can react in a meaningful way to control the rice pest S. mauritia by inducing cellular apoptosis in ovarian cells, which may cause adult females to become sterile.

HIV-1 mutants expressing B cell clonogenic matrix protein p17 variants are increasing their prevalence worldwide

AIDS-defining cancers declined after combined antiretroviral therapy (cART) introduction, but lymphomas are still elevated in HIV type 1 (HIV-1)-infected patients. In particular, non-Hodgkin's lymphomas (NHLs) represent the majority of all AIDS-defining cancers and are the most frequent cause of death in these patients. We have recently demonstrated that amino acid (aa) insertions at the HIV-1 matrix protein p17 COOH-terminal region cause protein destabilization, leading to conformational changes. Misfolded p17 variants (vp17s) strongly impact clonogenic B cell growth properties that may contribute to B cell lymphomagenesis as suggested by the significantly higher frequency of detection of vp17s with COOH-terminal aa insertions in plasma of HIV-1-infected patients with NHL. Here, we expand our previous observations by assessing the prevalence of vp17s in large retrospective cohorts of patients with and without lymphoma. We confirm the significantly higher prevalence of vp17s in lymphoma patients than in HIV-1-infected individuals without lymphoma. Analysis of 3,990 sequences deposited between 1985 and 2017 allowed us to highlight a worldwide increasing prevalence of HIV-1 mutants expressing vp17s over time. Since genomic surveillance uncovered a cluster of HIV-1 expressing a B cell clonogenic vp17 dated from 2011 to 2019, we conclude that aa insertions can be fixed in HIV-1 and that mutant viruses displaying B cell clonogenic vp17s are actively spreading.

Overexpression of MCAM induced by SMYD2-H3K36me2 in breast cancer stem cell properties.

Melanoma cell adhesion molecule (MCAM) is highly expressed in various malignancies. However, studies on the effects of MCAM on stemness of cancer stem cells are limited. Here, we aimed to explore the relationship between MCAM and stem cell phenotype in breast cancer (BC). We analyzed the genes differentially expressed in BC from the oncomine database, followed by TCGA-BRCA database validation. We then used gene set enrichment analysis to analyze the signaling pathways enriched to the relevant genes, followed by loss-of-function experiments to analyze the role of MCAM in the growth of BC cells and the maintenance of stem cell properties. We analyzed the cause for the MCAM overexpression using ChIP-seq and clarified the upstream mechanism by constructing SE-Deleted cells. Finally, the role of SMYD2 in the growth of BC cells and the maintenance of stem cell properties were verified by rescue experiments. MCAM was significantly overexpressed in BC, which predicted somber prognosis in patients. Knockdown of MCAM drastically hindered the growth and metastasis of BC cells in vitro and in vivo. Subsequently, the MCAM promoter was observed to have significant H3K36me2 modification and that SMYD2 could significantly promote the expression of MCAM. In addition, further overexpression of SMYD2 in cells with MCAM knockdown increased MCAM expression and promoted the growth as well as stemness of BC cells. SMYD2 can elevate the expression of MCAM by promoting its H3K36me2 modification, which in turn expedites the growth and stem cell properties of BC cells.

Purinergic and Adenosinergic Signaling in Pancreatobiliary Diseases.

Adenosine 5'-triphosphate (ATP), other nucleotides, and the nucleoside analogue, adenosine, all have the capacity to modulate cellular signaling pathways. The cellular processes linked to extracellular purinergic signaling are crucial in the initiation, evolution, and resolution of inflammation. Injured or dying cells in the pancreatobiliary tract secrete or release ATP, which results in sustained purinergic signaling mediated through ATP type-2 purinergic receptors (P2R). This process can result in chronic inflammation, fibrosis, and tumor development. In contrast, signaling via the extracellular nucleoside derivative adenosine via type-1 purinergic receptors (P1R) is largely anti-inflammatory, promoting healing. Failure to resolve inflammation, as in the context of primary sclerosing cholangitis or chronic pancreatitis, is a risk factor for parenchymal and end-organ scarring with the associated risk of pancreatobiliary malignancies. Emerging immunotherapeutic strategies suggest that targeting purinergic and adenosinergic signaling can impact the growth and invasive properties of cancer cells, potentiate anti-tumor immunity, and also block angiogenesis. In this review, we dissect out implications of disordered purinergic responses in scar formation, end-organ injury, and in tumor development. We conclude by addressing promising opportunities for modulation of purinergic/adenosinergic signaling in the prevention and treatment of pancreatobiliary diseases, inclusive of cancer.

Mammalian HEMK1 methylates glutamine residue of the GGQ motif of mitochondrial release factors

Despite limited reports on glutamine methylation, methylated glutamine is found to be highly conserved in a "GGQ" motif in both prokaryotes and eukaryotes. In bacteria, glutamine methylation of peptide chain release factors 1/2 (RF1/2) by the enzyme PrmC is essential for translational termination and transcript recycling. Two PrmC homologs, HEMK1 and HEMK2, are found in mammals. In contrast to those of HEMK2, the biochemical properties and biological significance of HEMK1 remain largely unknown. In this study, we demonstrated that HEMK1 is an active methyltransferase for the glutamine residue of the GGQ motif of all four putative mitochondrial release factors (mtRFs)—MTRF1, MTRF1L, MRPL58, and MTRFR. In HEMK1-deficient HeLa cells, GGQ motif glutamine methylation was absent in all the mtRFs. We examined cell growth and mitochondrial properties, but disruption of the HEMK1 gene had no considerable impact on the overall cell growth, mtDNA copy number, mitochondrial membrane potential, and mitochondrial protein synthesis under regular culture condition with glucose as a carbon source. Furthermore, cell growth potential of HEMK1 KO cells was still maintained in the respiratory condition with galactose medium. Our results suggest that HEMK1 mediates the GGQ methylation of all four mtRFs in human cells; however, this specific modification seems mostly dispensable in cell growth and mitochondrial protein homeostasis at least for HeLa cells under fermentative culture condition.

Controlling the Formation of Foams in Broth to Promote the Co-Production of Microbial Oil and Exopolysaccharide in Fed-Batch Fermentation

A large amount of foam is generated in the production of microbial oil and exopolysaccharide (EPS) by Sporidiobolus pararoseus JD-2, which causes low efficiency in fermentation. In this study, we aimed to reduce the negative effects of foams on the co-production of oil and EPS by controlling the formation of foams in broth. As we have found, the formation of foams is positively associated with cell growth state, air entrapment, and properties of broth. The efficient foam-control method of adding 0.03% (v/v) of the emulsified polyoxyethylene polyoxypropylene pentaerythritol ether (PPE) and feeding corn steep liquor (CSL) at 8–24 h with speed of 0.02 L/h considerably improved the fermentation performance of S. pararoseus JD-2, and significantly increased the oil and EPS concentrations by 8.7% and 12.9%, respectively. The biomass, oil, and EPS concentrations were further increased using a foam backflow device combined with adding 0.03% (v/v) of the emulsified PPE and feeding CSL at 8–24 h, which reached to 62.3 ± 1.8 g/L, 31.2 ± 0.8 g/L, and 10.9 ± 0.4 g/L, respectively. The effective strategy for controlling the formation of foams in fermentation broth reported here could be used as a technical reference for producing frothing products in fed-batch fermentation.

Targeted Knockout of the dhfr, glul, bak1, and bax Genes by the Multiplex Genome Editing in CHO Cells.

The Chinese hamster ovary cell line CHO is widely used for biopharmaceutical production. Genome editing makes it possible to improve the growth properties of cells, their auxotrophy, and the functioning of the apoptosis and autophagy induction systems. Simultaneous editing of multiple genes makes it possible to obtain a cell line with the required genotype faster than several consecutive rounds of genomic knockout, but the probability of success is lower. Simultaneous editing of the dhfr, glul, bak1, and bax genes in the CHO S cells genome yielded 24 clones with signs of auxotrophy for thymidine and glutamine. Five ofthem turned out to be dhfr+/-, all five contained a knockout of one or two glul alleles. In one clone, 7 out of 8 target alleles were inactivated by a frameshift, and the second dhfr allele was partially inactivated by insertion of the GAA triplet, which reduced the enzyme activity 2.5 times. The probability of simultaneous knockout of both dhfr alleles increased to 50% when the genome was edited with a pair of guide RNAs directed to one exon of the dhfr gene.

Evaluation of osteoblastic cell behavior upon culture on titanium substrates photo-functionalized by vacuum ultra-violet treatment

Photo-functionalization of titanium orthopedic/prosthetic implants using ultraviolet illumination is known to improve osteogenesis. Therefore, in this study, we aimed to examine the influence of vacuum ultraviolet (VUV)-treated titanium surfaces on osteoblast cell adhesion, activity, and differentiation. Osteoblastic cells were cultured on titanium substrates treated with various VUV treatment conditions (0, 6.2, 18.7, and 37.4 J/cm2) and their behavior was evaluated. The results revealed that cell adhesion was increased whereas cell activity and differentiation ability were decreased upon cell culture on VUV-treated substrates. In particular, cell activity and differentiation ability were dramatically suppressed with 18.7 J/cm2 VUV irradiation. Within the limitations of this cell-based experiment, we clarified the VUV treatment conditions in which cell adhesion was improved but cell activity and differentiation ability were suppressed. These results indicate that VUV-treatment can be used to influence cell growth properties and can be used to accelerate or suppress cell differentiation on implant substrates.

Abstract 12544: A miRNA Cassette for Reprogramming Vascular Smooth Muscle Cells Into Endothelial Cells

Introduction: Vascular smooth muscle cells (SMCs) and endothelial cells (ECs) exhibit unique miRNA profiles, which regulate their overall function. However, whether differentially expressed miRNAs can be used for transcriptional reprogramming of SMCs into ECs is unknown. MiRNA profiling of SMCs and ECs revealed differential regulation of miR-143-3p, miR-145-3p, miR-146a-5p, and miR-181b-5p as a set of candidates for transdifferentiation. Methods: Human coronary SMCs were transfected with miR-143-3p and miR-145-3p inhibitors alongside miR-146a-5p and miR-181b-5p mimics and selectively expanded. Induced endothelial cells (iECs) were then assessed for their transcriptional, protein expression, and functional similarities to ECs (Fig. 1). Results: Principal component and gene set enrichment analyses determined that iECs are transcriptionally clustered closer to ECs than SMCs in whole transcriptome profiling and selected EC and SMC gene sets. Top pathways included ‘blood vessel morphogenesis’, ‘regulation of angiogenesis’ and ‘Notch, VEGF, integrin signaling’. Transcriptome data was validated via PCR and Western blot analyses as iECs exhibited robust upregulation in EC markers CD31, VE-Cadherin, and VEGFR2 and downregulation of SMC markers MHY11, Vimentin, and αSMA. The similarity of iECs to ECs was functional as iECs demonstrated oxLDL and Ac-LDL uptake, tube formation in Matrigel™ assays, eNOS phosphorylation in response to VEGF, and iEC migratory and cell growth properties were similar to human umbilical vein ECs in the presence of VEGF. A hindlimb ischemia model validated iEC similarity to ECs as injected iECs restored blood flow and significantly reduced limb necrosis similar to ECs. Conclusion: A miRNA cassette consisting of miR-143-3p and miR-145-3p inhibitors and miR-146a-5p and miR-181b-5p mimics transformed SMCs into endothelial-like cells and show promise as a potential novel regenerative strategy for endothelial cells.

Engineered Biocompatible and Stable Dipeptide Hydrogel with Tunable Mechanical and Cell Growth Properties to Embolden Neuroglial Cell Growth

Engineered Biocompatible and Stable Dipeptide Hydrogel with Tunable Mechanical and Cell Growth Properties to Embolden Neuroglial Cell Growth

Surface Integrity and Biological Response of Ti-Alloy Implants after Surface Modification

The objective of present review is to critically analyze the induced surface characteristics and in-vitro & in-vivo analysis of Titanium and Ti-alloys based implants after surface modification through various techniques. The modified surfaces have nano-pores, micro cracks and recast layers which affects the properties of cell adhesion, osseointegration, cell growth and hemocompatibilty. EDM/WEDM have great potential for surface modifications of solid Ti and Ti-alloys for their use in the orthopaedic implants having better outcomes than other techniques. However, the stress shielding between the implants and bone is a major challenge in tissue engineering due to differences in their elastic moduli. The porous and perforated implants fabricated using micro-drilling, SEBM and SLM provide favorable solution to this problem. Therefore, through this literature review, the influence of adavance manufacturing processes on the biocompatibility, corrosion resistance and mechanical properties of Ti and Ti based alloys through in-vivo and in-vitro studies has been discused and reviewed.

Chitosan/PVA Based Membranes Processed by Gamma Radiation as Scaffolding Materials for Skin Regeneration.

Some of the current strategies for the development of scaffolding materials capable of inducing tissue regeneration have been based on the use of polymeric biomaterials. Chitosan, in particular, due to its recognized biological activity has been used in a number of biomedical applications. Aiming the development of chitosan-based membranes with improved cell adhesion and growth properties to be used as skin scaffolds allowing functional tissue replacement, different formulations with chitosan of different molecular weight, poly (vinyl alcohol) and gelatin, were evaluated. To meet the goal of getting ready-to-use scaffolds assuring membranes’ required properties and sterilization, preparation methodology included a lyophilization procedure followed by a final gamma irradiation step. Two radiation dose values were tested. Samples were characterized by TGA, FTIR, and SEM techniques. Their hydrophilic properties, in vitro stability, and biocompatibility were also evaluated. Results show that all membranes present a sponge-type inner structure. Chitosan of low molecular weight and the introduction of gelatin are more favorable to cellular growth leading to an improvement on cells’ morphology and cytoskeletal organization, giving a good perspective to the use of these membranes as potential skin scaffolds.

Circular RNA circSETD3 hampers cell growth, migration, and stem cell properties in bladder cancer through sponging miR-641 to upregulate PTEN

ABSTRACT Bladder cancer (BLCA) is a common malignant urothelial cancer in the world. Although circular RNAs (circRNAs) involve in regulating BLCA progression, the role of a novel circular RNA circSETD3 in regulating BLCA pathogenesis has not been studied. The expression of circSETD3, miR-641, PTEN mRNA in BLCA tissues and cell lines were measured using RT-qPCR. The gain-of-function experiments were performed in vitro and in vivo to detect the effects of circSETD3 on cell proliferation, migration, EMT, and stemness maintenance. Besides, rescue experiments were performed to demonstrate the regulatory mechanism of circSETD3/miR-641/PTEN in BLCA cell malignant phenotypes in vitro. CircSETD3 was remarkably downregulated in the cancerous clinical tissues and cell lines, in contrast with their normal counterparts, and circSETD3 tended to be deficient in BLCA patients with larger tumor size, advanced clinical stages, positive lymph metastasis and worse prognosis. In addition, circular isoforms of circSETD3 were more resistant to RNase R+ and actinomycetes D treatment compared to their linear isoforms, and circSETD3 mainly distributed in the cytoplasm of the BLCA cells. Further gain-of-function experiments showed that circSETD3 acted as a tumor suppressor to suppress BLCA cell proliferation, migration, EMT and stemness, and the underlying mechanisms had also been elucidated. Mechanistically, circSETD3 sponged miR-641 to upregulate PTEN, resulting in the blockage of BLCA progression. Our findings indicated that circSETD3 acted as a vital tumor suppressor in BLCA via regulating the miR-641/PTEN axis.

Role of CD10 expression in endometriosis-associated mesenchymal stem cells on the progression of endometriosis-associated carcinoma.

5561 Background: Endometriosis-associated carcinomas (EACs) such as ovarian clear cell cancer (OCCC) are rare, aggressive, chemo-resistant malignancies. While endometriosis is a known chronic inflammatory condition, the molecular mechanisms for the malignant transformation of endometriosis is unknown. Mesenchymal stem cells (MSC) are a critical component of the ovarian cancer microenvironment. Cancer cells reprogram MSCs to form carcinoma-associated MSCs (CAMSCs), which promote cancer growth, chemotherapy resistance, and metastases. MSCs are also found within the endometriotic microenvironment. CD10, a surface protein expressed by endometrial stromal cells, is also expressed on endometriosis-associated MSCs (enMSCs). Preliminary data demonstrate CD10 expression is lost in a subset of enMSCs and this loss is correlated with the acquisition of tumor-promoting properties. We hypothesized that the CD10 negative subset of enMSCs behave similarly to CAMSCs and support the growth of OCCC. Methods: EnMSCs were isolated from primary human benign endometriosis deposits involving the ovary or fallopian tubes. Flow cytometry was used to measure surface CD10 expression. We investigated the role of low CD10 enMSCs versus high CD10 enMSCs on OCCC tumor cell growth, chemotherapy resistance and stem-like cell properties in vitro and tumor cell engraftment, growth, and metastases in vivo. Luciferase-expressing OCCC cells were (1) used alone, (2) mixed with low CD10 enMSCs, or (3) mixed with high CD10 enMSCs and injected orthotopically into the ovarian bursa of NSG mice. In vivo imaging system was used to follow tumor progression and metastasis. Results: Our results demonstrated that enMSCs have variable CD10 expression. EnMSCs with low CD10 expression significantly enhanced OCCC proliferation, resistance to cisplatin, and sphere formation compared to OCCC alone. In contrast, high CD10 expressing enMSCs significantly reduce OCCC proliferation and sphere formation. Interestingly, low CD10 enMSCs selectively enhanced OCCC cell growth and had no effect on high grade serious ovarian cancer cell growth. Moreover, a reduction of CD10 expression was observed over time when high CD10 enMSCs were co-cultured with OCCC cells. Our results also showed enhanced tumor engraftment when OCCC cells were co-injected with low CD10 enMSCs to 100% one week post-injection, compared to 40% with OCCC and high CD10 enMSCs and 60% with OCCC alone. Further, mice co-injected with low CD10 enMSCs demonstrated increased metastasis and decreased survival compared to mice co-injected with high CD10 enMSCs. Conclusions: Our results indicate there is a sub population of enMSCs, marked by decreased CD10 expression, which selectively enhances OCCC growth. This highlights the existence of a tumor-promoting stromal cell within endometriosis which may be critical to the formation and propagation of EACs.

WITHDRAWN: ERK-CREB pathway is involved in HSPB8-mediated glioma cell growth and metastatic properties

To investigate the mechanism of HSPB8 (heat shock protein beta-8) in the growth and metastatic properties of glioma cells. HSPB8 expression in glioma tissue and cell was detected via Western blotting. Then, glioma U87 and U251 cell lines were divided into Mock group, Control siRNA group, HSPB8 siRNA-1 group and HSPB8 siRNA-2 group. Cell proliferation was detected using MTT assay, while its invasion, migration and apoptosis were determined by Transwell, wound-healing and flow cytometry, respectively. The expression of HSPB8 and ERK-CREB pathway-related molecules were also measured by Western blotting. Xenograft models were constructed on nude mice, and accordingly, the growth curve of subcutaneous xenograft was prepared. In glioma tissues, HSPB8 expression was upregulated with the increasing grade of glioma. Besides, glioma cells in the HSPB8 siRNA-1 group and HSPB8 siRNA-2 group manifested the significant enhancement in apoptotic rates and reductions in its proliferation, migration and invasion compared to those in the Mock group, meanwhile, the expression of HSPB8, p-ERK1/2/ERK1/2 and p-CREB/CREB were downregulated. On the other hand, the tumor growth in the nude mice of Ad-HSPB8 shRNA-1 group and Ad-HSPB8 shRNA-2 group was retarded significantly, with an acute decrease in the tumor weight. Silencing HSPB8 can inhibit the malignant features, while facilitate the apoptosis of glioma cells, with inactivation of ERK-CREB pathway.

Water jet-assisted lipoaspiration and Sepax cell separation system for the isolation of adipose stem cells with high adipogenic potential

Water jet-assisted liposuction has gained popularity due to favourable fat grafting outcomes. In this study, we compared stem cells obtained from fat isolated with manual or the water jet-assisted procedure. Liposuction of abdominal fat was performed using the two methods on each donor (n = 10). Aspirate samples were collagenase digested and the isolated cells seeded in vitro prior to proliferation, adipogenic differentiation and angiogenic activity analyses. Cells from either procedure proliferated at similar rates and exhibited a similar colony-forming ability. The cells expressed stem cell markers CD73, CD90 and CD105. In the water jet cell preparations, there were higher numbers of cells expressing CD146. Robust adipogenic differentiation was observed in cultures expanded from both manual and water jet lipoaspirates. Gene analysis showed higher expression of the adipocyte markers aP2 and GLUT4 in the adipocyte-differentiated water jet cell preparations, and ELISA indicated increased secretion of adiponectin from these cells. Both cell groups expressed vasculogenic factors and the water jet cells promoted the highest levels of in vitro angiogenesis. Given these positive results, we further characterised the water jet cells when prepared using an automated closed cell processing unit, the Sepax-2 system (Cytiva). The growth and stem cell properties of the Sepax-processed cells were similar to the standard centrifugation protocol, but there was evidence for greater adipogenic differentiation in the Sepax-processed cells. Water jet lipoaspirates yield cells with high adipogenic potential and angiogenic activity, which may be beneficial for use in cell-assisted lipotransfers.

Evaluation of the cytocompatibility of methacrylate resin-based root canal sealers with osteoblast-like cells.

This study compared the cytocompatibilities of three methacrylate resin-based root canal sealers [MetaSEAL Soft (MSS), Hybrid Root SEAL (HRS), and Superbond Sealer (SBS)] in either freshly mixed or set conditions using the Kusa A1 osteoblastic cell line. The three sealers and an epoxy resin-based sealer (AH Plus; AHP) were extracted in culture medium; cell growth and osteogenic properties were analyzed. Cell adhesion on set sealers was analyzed with scanning electron microscopy. The respective extents of cell growth were as follows in freshly mixed and set sealer extracts: SBS>MSS>AHP>HRS and SBS=AHP>MSS>HRS. Light irradiation of MSS and HRS increased the cell growth of set sealer extracts. Set SBS, MSS, and AHP did not alter expression of osteogenic genes or formation of mineralized nodules. Attached cells were observed only on SBS. In conclusion, the four sealers exhibited varying degrees of compatibility to osteoblasts; SBS and HRS were the most and least compatible, respectively.