1,721,008 research outputs found
Mesenchymal stem cells and nano-structured surfaces
No full textMesenchymal stem cells (MSCs) represent multipotent stromal cells that can differentiate into a variety of cell types, including osteoblasts (bone cells), chondrocytes (cartilage cells), and adipocytes (fat cells). Their multi-potency provides a great promise as a cell source for tissue engineering and cell-based therapy for many diseases, particularly bone diseases and bone formation. To be able to direct and modulate the differentiation of MSCs into the desired cell types in situ in the tissue, nanotechnology is introduced and used to facilitate or promote cell growth and differentiation. These nano-materials can provide a fine structure and tuneable surface in nanoscales to help the cell adhesion and promote the cell growth and differentiation of MSCs. This could be a dominant direction in future for stem cells based therapy or tissue engineering for various diseases. Therefore, the isolation, manipulation, and differentiation of MSCs are very important steps to make meaningful use of MSCs for disease treatments. In this chapter, we have described a method of isolating MSC from human bone marrow, and how to culture and differentiate them in vitro. We have also provided research methods on how to use MSCs in an in vitro model and how to observe MSC biological response on the surface of nano-scaled materials
Isolation of cancer stem cells from squamous cell carcinoma
No full textDifferent cancer stem cell (CSC) populations can be found in many types of cancer, including squamous cell carcinoma (SSC). Diverse reports showed that CSC play a crucial role in the relapse of different types of cancer. CSC sustains tumor growth due to their capacity to self-renew and their potential to initiate secondary tumors with metastatic cancer features. Therefore, the development of methods for the isolation of CSC is a key step to explore the mechanisms underlying CSC maintenance. In this chapter, we provide a method for isolating CSC from cutaneous SSC using immunofluorescence labeling to allow the specific purification of CSC by fluorescence-activated cell sorting (FACS). This method is based on the use of CSC membrane markers, allowing as well the isolation CSC from different mouse strains.</p
Evaluation of Mitochondrial Phagy (Mitophagy) in Human Non-small Adenocarcinoma Tumor Cells
No full textAcknowledgments
SG conceptualized and designed the study. JA performed the experiments. SG and JA SCDR analyzed the data. JA and MC wrote the manuscript. SG, JA, SCDR, and MC reviewed and edited the manuscript. SG funded and supervised the experiments.
Funding
This study was funded by Canercare Manitoba Foundation. JA was supported by CIHR Vaneir PhD studentship. SCDR was supported with a Postdoctoral Fellowship sponsored by the Canadian Institutes of Health and Research (CIHR), FRN 176508. MC was supported by grant RYC2021-031003I funded by MICIU/AEI/10.13039/501100011033 and, by European Union NextGenerationEU/PRTR.Non–small cell lung cancer (NSCLC) is a predominant form of lung cancer characterized by its aggressive nature and high mortality rate, primarily due to late-stage diagnosis and metastatic spread. Recent studies underscore the pivotal role of mitophagy, a selective form of autophagy targeting damaged or superfluous mitochondria, in cancer biology, including NSCLC. Mitophagy regulation may influence cancer cell survival, proliferation, and metastasis by modulating mitochondrial quality and cellular energy homeostasis. Herein, we present a comprehensive methodology developed in our laboratory for the evaluation of mitophagy in NSCLC tumor cells. Utilizing a combination of immunoblotting, immunocytochemistry, and fluorescent microscopy, we detail the steps to quantify early and late mitophagy markers and mitochondrial dynamics. Our findings highlight the potential of targeting mitophagy pathways as a novel therapeutic strategy in NSCLC, offering insights into the complex interplay between mitochondrial dysfunction and tumor progression. This study not only sheds light on the significance of mitophagy in NSCLC but also establishes a foundational approach for its investigation, paving way for future research in this critical area of cancer biology.Canercare Manitoba FoundationCanadian Institutes of Health and Research (CIHR)Unión EuropeaMinisterio de Ciencia, Innovación y Universidades (España)Depto. de Bioquímica y Biología MolecularFac. de Ciencias BiológicasTRUEpu
Assessing autophagy flux in glioblastoma temozolomide resistant cells
No full textFunding:
M.C. is supported by grant RYC2021-031003I funded by MICIU/AEI/https://doi.org/10.13039/501100011033 and by European Union NextGenerationEU/PRTR.Autophagy is a critical cellular process involved in the degradation and recycling of cytoplasmic components, playing a dual role in cancer by either promoting cell survival or facilitating cell death. In glioblastoma (GB), autophagy has been implicated in resistance to the chemotherapeutic agent temozolomide (TMZ). This study presents a novel method to accurately measure autophagy flux in TMZ-resistant glioblastoma cells, combining advanced imaging techniques with biochemical assays. By quantifying key autophagy markers such as LC3-II and SQSTM1, our approach provides detailed insights into the dynamic processes of autophagosome formation and clearance under therapeutic stress. This method advances our understanding of autophagy in GB chemoresistance and has significant implications for the development of autophagy-targeted therapies. The ability to monitor and manipulate autophagy flux in real time offers a promising avenue for monitoring and understanding TMZ resistance and improving patient outcomes in glioblastoma treatment.European CommissionMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de InvestigaciónDepto. de Bioquímica y Biología MolecularFac. de Ciencias BiológicasTRUEpu
Development of biological tools to study claudins in the male reproductive tract
No full textIt is estimated that between 12 and 15% of couples are infertile. More than half of these are related to problems associated with male reproductive dysfunction. Of those, 40% occur from idiopathic or unexplained causes. While spermatozoa are formed in the testis, testicular spermatozoa are immature and cannot swim or fertilize. These critical functions are acquired as spermatozoa transit through the epididymis in the specific luminal environment created in part by the tight junctions of the blood-epididymis barrier. To understand the normal and pathological conditions attributable to human and animal epididymal function, we have needed to develop biological tools to characterize the physiological, cellular, and molecular functions of tight junctions and claudins (Cldns) in the epididymis. We have shown that by developing epididymal cell lines we have gained valuable insight into the functions of epididymal Cldns, the regulation of the Cldn1 gene and how these can be mistargeted in infertile men. Here we describe some of the techniques that have been used to address these critical aspects of epididymal Cldns.[on SciFinder (R)]</br
In vivo optical imaging of ischemic blood\u2013brain barrier disruption
No full textPeer reviewed: YesNRC publication: Ye
Lineage selection for generation and amplification of neural precursor cells
No full textEmbryonic stem (ES) cells are derived from the epiblast of mouse blastocyst. They can repopulate all cell lineages in vivo and can differentiate into a wide variety of cell types in vitro during embryoid body (EB) formation (1). ES cells have been shown to generate both neurones and glial cells (2,3). During the course of ES cell differentiation, neural precursors that express nestin and/or sox1 and sox2 appear first, these are followed by βtubulin 3 and neurofilament-expressing neurons and, subsequently, glial fibrillary-acid protein (GFAP) or O4 positive glial cells (4–8). These results suggest that ES cell-derived neural system can be used for experimental dissection of various aspects of mammalian neural development. If extended to humans, in vitro-generated neural cells could also be used as source for transplantation-based cell therapy
The Male Stem Cell Niche: Insights from Drosophila and Mammalian Model Systems
No full textStem cells are indispensable for multicellular development since they provide the cells that built up our bodies and renew damaged cells during adult life. Specialized inputs for proper stem cell function lay in a local tissue microenvironment, the stem cell niche that homes them and regulates the balance between stem cell renewal and differentiation. Spermatogenesis is a classical adult stem cell system in which germline stem cells generate the male gametes that transmit genetic information to the progeny of the organism. A common theme in spermatogenesis across various species is that male stem cell niche and testis morphogenesis relies on intrinsic germline stem cell factors but also on the physical contact and diffusible signals exchanged between the germline and the juxtaposed somatic cell populations. This book chapter addresses the current state of the art of the male stem cell niche in Drosophila and mammalian testes. We review well-established and recent findings on male stem cell niche architecture, and on how the niche and germline cross-talk shapes spermatogenesis and promotes normal sperm production, which is of outmost importance for all sexually reproductive organisms. Finally, we compare Drosophila and mammalian male stem cell niches, discuss the differences and similarities, and provide stimulating insights on the basic organizing and regulatory strategies adapted in each model organism
Adult Stromal (Skeletal, Mesenchymal) Stem Cells: Advances Towards Clinical Applications.
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