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Bio-medical Materials and Engineering 2021Merging stem cells with biomimetic materials represent an attractive approach to tissue engineering. The development of an alternative scaffold with the ability to mimic...
BACKGROUND
Merging stem cells with biomimetic materials represent an attractive approach to tissue engineering. The development of an alternative scaffold with the ability to mimic the extracellular matrix, and the 3D gradient preventing any alteration in cell metabolism or in their gene expression patterns, would have many medical applications.
OBJECTIVE
In this study, we introduced the use of RGD (Arg-Gly-Asp) bio-conjugated cotton to promote the growth and proliferation of mesenchymal stem cells (MSCs).
METHODS
We measured the expression of stem cell markers and adhesion markers with Q-PCR and analyzed the transcriptomic. The results obtained showed that the MSCs, when cultured with bio-conjugated cotton fibers, form aggregates around the fibers while proliferating. The seeded MSCs with cotton fibers proliferated in a similar fashion to the cells seeded on the monolayer (population doubling level 1.88 and 2.19 respectively).
RESULTS
The whole genome sequencing of cells adhering to these cotton fibers and cells adhering to the cell culture dish showed differently expressed genes and pathways in both populations. However, the expression of the stem cell markers (Oct4, cKit, CD105) and cell adhesion markers (CD29, HSPG2 and CD138), when examined with quantitative RT-PCR, was maintained in both cell populations.
CONCLUSION
These results clearly show the ability of the cotton fibers to promote MSCs growth and proliferation in a 3D structure mimicking the in vivo environment without losing their stem cell phenotype.
Topics: Cell Differentiation; Cell Proliferation; Cells, Cultured; Cotton Fiber; Mesenchymal Stem Cells; Oligopeptides; Tissue Scaffolds
PubMed: 33164919
DOI: 10.3233/BME-201115 -
Biomedicine & Pharmacotherapy =... Oct 2019Hypoxia has been suggested to be both beneficial and harmful to the proliferation of cardiomyocytes. This controversy remains unresolved, and the underlying mechanism by...
BACKGROUND
Hypoxia has been suggested to be both beneficial and harmful to the proliferation of cardiomyocytes. This controversy remains unresolved, and the underlying mechanism by which hypoxia exerts its effects remains unclear. We here hypothesize that cardiomyocyte developmental stage may play a role.
METHODS AND RESULTS
The embryonic ventricular myocyte cell line H9C2, primary isolated fetal cardiomyocytes, and neonatal cardiomyocytes were cultured with normal O (21% O) or under hypoxic conditions (10% O) for 7 days, and then harvested for Western blotting, qRT-PCR, and immunostaining. When cultured under hypoxic conditions, proliferating marker-Ki67, mRNA level, and the percentage of Ki67-positive cardiomyocytes were significantly lower in H9C2 and fetal cardiomyocytes but higher in neonatal cardiomyocytes. Consistently, the mRNA and protein levels and induced nuclear localization of yes associated protein 1(YAP1), one of the most important regulators of cardiomyocyte proliferation, were significantly lower in H9C2 and fetal cardiomyocytes but up-regulated in neonatal cardiomyocytes when treated with hypoxia. Compared to neonatal cardiomyocytes, there was a lower level of troponin T mRNA and protein expression in H9C2 and fetal cardiomyocytes. When H9C2 or fetal cardiomyocytes overexpressing troponin T in were cultured under hypoxic condition, their ability to proliferate increased.
CONCLUSIONS
The effect of hypoxia on the proliferation of cardiomyocyte is associated with their developmental stage. YAP1 expression is positively correlated with the change in cardiomyocyte proliferation in response to hypoxia. Developmental stage- specific sarcomere component troponin T may partly account for the underlying mechanism.
Topics: Animals; Animals, Newborn; Apoptosis Regulatory Proteins; Cell Hypoxia; Cell Line; Cell Proliferation; Cells, Cultured; Models, Biological; Myocytes, Cardiac; Rats; Troponin T; YAP-Signaling Proteins
PubMed: 31545287
DOI: 10.1016/j.biopha.2019.109391 -
Virchows Archiv : An International... Jun 2018Benign proliferations that mimic malignancies are commonly encountered during the course of assessment of small and fragmented endometrial samples. Although benign,... (Review)
Review
Benign proliferations that mimic malignancies are commonly encountered during the course of assessment of small and fragmented endometrial samples. Although benign, endometrial epithelial metaplasias often coexist with premalignant or malignant lesions causing diagnostic confusion. The difficulty with mucinous metaplasia lies in its distinction from atypical mucinous glandular proliferations and mucinous carcinomas, which are associated with significant interobserver variability. Papillary proliferation of the endometrium is commonly associated with hormonal drugs and endometrial polyps and is characterised by papillae with fibrovascular cores covered by epithelial cells without cytologic atypia. They are classified into simple or complex papillary proliferations depending on the architectural complexity and extent of proliferation. Complex papillary proliferations are associated with a high risk of concurrent or subsequent hyperplasia with atypia/carcinoma. Papillary proliferations may have coexisting epithelial metaplasias and, most commonly, mucinous metaplasia and syncytial papillary change. Those with striking mucinous metaplasia overlap morphologically with papillary mucinous metaplasia. The latter has been proposed as a precursor of endometrial mucinous carcinoma. Misinterpreting the Arias-Stella reaction as a malignant or premalignant lesion is more likely to occur if the pathologist is unaware that the patient is pregnant or on hormonal drugs. Endometrial hyperplasia with secretory changes may occasionally be difficult to distinguish from the torturous and crowded glands of a late secretory endometrium. Endometrial polyps may have abnormal features that can be misinterpreted as endometrial hyperplasia or Mullerian adenosarcoma. Awareness of these benign endometrial proliferations and their common association with hormonal medication or altered endogenous hormonal levels will help prevent the over-diagnosis of premalignant and malignant lesions.
Topics: Biopsy; Carcinoma, Endometrioid; Cell Proliferation; Endometrial Neoplasms; Endometrium; Female; Humans; Metaplasia
PubMed: 29445890
DOI: 10.1007/s00428-018-2314-4 -
Transplantation May 2022Activation of porcine endothelial cells (PECs) is the mechanistic centerpiece of xenograft rejection. This study sought to characterize the immuno-phenotype of human T...
BACKGROUND
Activation of porcine endothelial cells (PECs) is the mechanistic centerpiece of xenograft rejection. This study sought to characterize the immuno-phenotype of human T cells in response to PECs and to explore the immuno-modulation of B7 and mammalian target of rapamycin blockade of T cells and/or PECs during xeno-responses.
METHODS
Rapid memory T-cell (TM) responses to PECs were assessed by an intracellular cytokine staining. T-cell proliferation to PEC with or without belatacept or rapamycin was evaluated by a mixed lymphocyte-endothelial cell reaction (MLER). Additionally, rapamycin-pretreated PECs were used in MLER. Cell phenotypes were analyzed by flow cytometry.
RESULTS
Tumor necrosis factor-α/interferon-γ producers were detected in CD8+ cells stimulated by human endothelium but not PECs. MLER showed proliferation of CD4+ and CD8+ cells with predominantly memory subsets. Purified memory and naive cells proliferated following PEC stimulation with an increased frequency of TM in PEC-stimulated naive cells. Proliferating cells upregulated programmed cell death-1 (PD-1) and CD2 expression. Belatacept partially inhibited T-cell proliferation with reduced CD2 expression and frequency of the CD8+CD2highCD28- subset. Rapamycin dramatically inhibited PEC-induced T-cell proliferation, and rapamycin-preconditioned PECs failed to induce T-cell proliferation. PD-1 blockade did not restore T-cell proliferation to rapamycin-preconditioned PECs.
CONCLUSIONS
Humans lack rapid TM-mediated responses to PECs but induce T-cell proliferative responses characterized largely as TM with increasing CD2 and PD-1 expression. B7-CD28 and mammalian target of rapamycin blockade of T cells exhibit dramatic inhibitory effects in altering xeno-proliferating cells. Rapamycin alters PEC xeno-immunogenicity leading to inhibition of xeno-specific T-cell proliferation independent of PD-1-PD ligand interaction.
Topics: Abatacept; Animals; B7 Antigens; Cell Proliferation; Endothelial Cells; Humans; Mammals; Programmed Cell Death 1 Receptor; Sirolimus; Swine; TOR Serine-Threonine Kinases
PubMed: 34387242
DOI: 10.1097/TP.0000000000003920 -
Archives of Medical Science : AMS 2023To explore the function of interleukin 1α (IL-1α) in bladder cancer (BCa).
INTRODUCTION
To explore the function of interleukin 1α (IL-1α) in bladder cancer (BCa).
MATERIAL AND METHODS
Immunohistochemistry (IHC) was used to test the protein expression of IL-1α in BCa tissues. The relationship between IL-1α and clinical characteristics was analyzed by the Kaplan-Meier curve method. The gene and protein expression was tested by reverse transcription (RT-q-PCR) and western blot, respectively. Colony formation and MTT assays were used to detect the potential of proliferation , and scratch and transwell chamber assays were used to detect the potential of invasion . Markers of proliferation such as Ki-67 and proliferating cell nuclear antigen (PCNA) and markers of invasion such as MMP-2 and MMP-9 were detected by western blot. Xenograft study was used for the experiment.
RESULTS
We found that IL-1α was highly expressed in BCa patients while highly expressed IL-1α was significantly related to short overall survival and progression-free survival in BCa as well. Moreover, knockdown of IL-1α might inhibit the ability of cancer cells to proliferate and invade or migrate both and .
CONCLUSIONS
Our findings suggested that IL-1α might be a therapy target for BCa malignant progression.
PubMed: 36817666
DOI: 10.5114/aoms.2020.100677 -
Biology Aug 2023Although microglia exist as a minor glial cell type in the normal state of the brain, they increase in number in response to various disorders and insults. However, it... (Review)
Review
Although microglia exist as a minor glial cell type in the normal state of the brain, they increase in number in response to various disorders and insults. However, it remains unclear whether microglia proliferate in the affected area, and the mechanism of the proliferation has long attracted the attention of researchers. We analyzed microglial mitosis using a facial nerve transection model in which the blood-brain barrier is left unimpaired when the nerves are axotomized. Our results showed that the levels of macrophage colony-stimulating factor (M-CSF), cFms (the receptor for M-CSF), cyclin A/D, and proliferating cell nuclear antigen (PCNA) were increased in microglia in the axotomized facial nucleus (axotFN). In vitro experiments revealed that M-CSF induced cFms, cyclin A/D, and PCNA in microglia, suggesting that microglia proliferate in response to M-CSF in vivo. In addition, M-CSF caused the activation of c-Jun N-terminal kinase (JNK) and p38, and the specific inhibitors of JNK and p38 arrested the microglial mitosis. JNK and p38 were shown to play roles in the induction of cyclins/PCNA and cFms, respectively. cFms was suggested to be induced through a signaling cascade of p38-mitogen- and stress-activated kinase-1 (MSK1)-cAMP-responsive element binding protein (CREB) and/or p38-activating transcription factor 2 (ATF2). Microglia proliferating in the axotFN are anticipated to serve as neuroprotective cells by supplying neurotrophic factors and/or scavenging excite toxins and reactive oxygen radicals.
PubMed: 37627005
DOI: 10.3390/biology12081121 -
PeerJ 2020Bryozoans are small benthic colonial animals; their colonies consist of zooids which are composed of a cystid and polypide. According to morphological and molecular...
Bryozoans are small benthic colonial animals; their colonies consist of zooids which are composed of a cystid and polypide. According to morphological and molecular data, three classes of bryozoans are recognized: Phylactolaemata, Gymnolaemata and Stenolaemata. Bryozoans are active suspension feeders and their feeding apparatus, the lophophore, is fringed with a single row of ciliated tentacles. In gymnolaemates, the lophophore is bell-shaped and its tentacles may be equal in length (equitentacled lophophores) or some tentacles may be longer than others (obliquely truncated lophophores). In encrusting colonies, polypides with obliquely truncated lophophores usually border specific sites of excurrent water outlets (colony periphery and chimneys) where depleted water has to be removed. It is known that during colony astogeny, colony-wide water currents rearrange: new chimneys are formed and/or location of the chimneys within a given colony changes with time. Such rearrangement requires remodeling of the lophophore shape and lengthening of some tentacles in polypides surrounding water outlets. However, proliferating activity has not been described for bryozoans. Here, we compared the distribution of S-phase and mitotic cells in young and adult polypides in three species of Gymnolaemata. We tested the hypothesis that tentacle growth/elongation is intercalary and cell proliferation takes place somewhere at the lophophore base because such pattern does not interfere with the feeding process. We also present a detailed description of ultrastructure of two parts of the lophophore base: the oral region and ciliated pits, and uncover the possible function of the latter. The presence of stem cells within the ciliated pits and the oral region of polypides provide evidence that both sites participate in tentacle elongation. This confirms the suggested hypothesis about intercalary tentacle growth which provides a potential to alter a lophophore shape in adult polypides according to rearrangement of colony wide water currents during colony astogeny. For the first time deuterosome-like structures were revealed during kinetosome biogenesis in the prospective multiciliated epithelial cells in invertebrates. Tentacle regeneration experiments in demonstrated that among all epidermal cell types, only non-ciliated cells at the abfrontal tentacle surface are responsible for wound healing. Ciliated cells on the frontal and lateral tentacle surfaces are specialized and unable to proliferate, not even under wound healing. Tentacle regeneration in is very slow and similar to the morphallaxis type. We suggest that damaged tentacles recover their length by a mechanism similar to normal growth, powered by proliferation of cells both within ciliated pits and the oral region.
PubMed: 32523809
DOI: 10.7717/peerj.9179 -
Frontiers in Immunology 2018Inosine monophosphate dehydrogenase (IMPDH) catalyzes the conversion of IMP to xanthosine monophosphate, the rate-limiting step in guanosine monophosphate (GMP)...
Inosine monophosphate dehydrogenase (IMPDH) catalyzes the conversion of IMP to xanthosine monophosphate, the rate-limiting step in guanosine monophosphate (GMP) synthesis. In cultured cells, IMPDH polymerizes into micron-scale filamentous structures when GMP synthesis is inhibited by depletion of purine precursors or by various drugs, including mycophenolic acid, ribavirin, and methotrexate. IMPDH filaments also spontaneously form in undifferentiated mouse embryonic stem cells and induced pluripotent stem cells, hinting they might function in various highly proliferative cell types. Therefore, we investigated IMPDH filament formation in human and murine T cells, which rely heavily on guanine nucleotide synthesis to rapidly proliferate in response to antigenic challenge. We discovered extensive IMPDH filament formation in mature T cells, B cells, and other proliferating splenocytes of normal, adult B6 mice. Both cortical and medullary thymocytes in young and old mice also showed considerable assembly of IMPDH filaments. We then stimulated primary human peripheral blood mononuclear cells with T cell mitogens phytohemagglutinin (PHA), concanavalin A (ConA), or antibodies to CD3 and CD28 for 72 h. We detected IMPDH filaments in 40-60% of T cells after activation compared to 0-10% of unstimulated T cells. Staining of activated T cells for the proliferation marker Ki-67 also showed an association between IMPDH filament formation and proliferation. Additionally, we transferred ovalbumin-specific CD4 T cells from B6.OT-II mice into B6.Ly5a recipient mice, challenged these mice with ovalbumin, and harvested spleens 6 days later. In these spleens, we identified abundant IMPDH filaments in transferred T cells by immunofluorescence, indicating that IMPDH also polymerizes during antigen-specific T cell activation. Overall, our data indicate that IMPDH filament formation is a novel aspect of T cell activation and proliferation, and that filaments might be useful morphological markers for T cell activation. The data also suggest that IMPDH filament formation could be occurring in a variety of proliferating cell types throughout the body. We propose that T cell activation will be a valuable model for future experiments probing the molecular mechanisms that drive IMPDH polymerization, as well as how IMPDH filament formation affects cell function.
Topics: Aging; Animals; B-Lymphocytes; Cell Proliferation; Female; Humans; IMP Dehydrogenase; Lymphocyte Activation; Male; Mice; Protein Multimerization; T-Lymphocytes
PubMed: 30555474
DOI: 10.3389/fimmu.2018.02789 -
Scientific Reports Sep 2021The gold nanorods (GNRs) embedded alginate-chitosan (scaffold), which was designed and fabricated to produce efficient handling of the cell proliferations. Scaffold...
The gold nanorods (GNRs) embedded alginate-chitosan (scaffold), which was designed and fabricated to produce efficient handling of the cell proliferations. Scaffold embedded GNR (SGNR) and NIR (near infrared) irradiations are developing into an interesting medical prognosis tool for rabbit chondrocyte (RC) proliferation. SGNR contained a pattern of uniform pores. Biocompatibility and cellular proliferation achieved by disclosures to NIR irradiations, providing high cell survival. SGNR and NIR irradiations could produce mechanical and biochemical cues for regulating RCs proliferations. To determine the thermal stress, it exposed RCs to 39-42 °C for 0-240 min at the start point of the cell culture cycle. It produced photothermal stress in cellular surrounding (cells located adjacent to and within scaffold) and it deals with the proliferation behavior of RC. All the processes were modeled with experimental criteria and time evolution process. Our system could help the cell proliferation by generating heat for cells. Hence, the present strategy could be implemented for supporting cell therapeutics after transplantation. This implementation would open new design techniques for integrating the interfaces between NIR irradiated and non-irradiated tissues.
Topics: Animals; Cell Proliferation; Cell Survival; Cells, Cultured; Chondrocytes; Gold; Nanotubes; Phototherapy; Rabbits
PubMed: 34588578
DOI: 10.1038/s41598-021-98929-2 -
Clinical & Translational Immunology 2019γδ T cells are fascinating cells that bridge the innate and adaptive immune systems. They have long been known to proliferate rapidly following infection; however, the... (Review)
Review
γδ T cells are fascinating cells that bridge the innate and adaptive immune systems. They have long been known to proliferate rapidly following infection; however, the identity of the specific γδ T cell subsets proliferating and the role of this expansion in protection from disease have only been explored more recently. Several recent studies have investigated γδ T-cell responses to vaccines targeting infections such as , and influenza, and studies in animal models have provided further insight into the association of these responses with improved clinical outcomes. In this review, we examine the evidence for a role for γδ T cells in vaccine-induced protection against various bacterial, protozoan and viral infections. We further discuss results suggesting potential mechanisms for protection, including cytokine-mediated direct and indirect killing of infected cells, and highlight remaining open questions in the field. Finally, building on current efforts to integrate strategies targeting γδ T cells into immunotherapies for cancer, we discuss potential approaches to improve vaccines for infectious diseases by inducing γδ T-cell activation and cytotoxicity.
PubMed: 31485329
DOI: 10.1002/cti2.1072