scholarly journals Menstrual Fluid Factors Mediate Endometrial Repair

2021 ◽  
Vol 3 ◽  
Author(s):  
Lois A. Salamonsen
Keyword(s):  
Ex Vivo ◽  
Endometrial Thickness ◽  
Live Cells ◽  
Cellular Debris ◽  
Reproductive Life ◽  
Keratinocyte Cell ◽  
Culture Models ◽  
Cellular Components ◽  
Menstrual Fluid

Menstruation is a process whereby the outer functionalis layer of the endometrium is shed each month in response to falling progesterone and estrogen levels in a non-conception cycle. Simultaneously with the tissue breakdown, the surface is re-epithelialized, protecting the wound from infection. Once menstruation is complete and estrogen levels start to rise, regeneration progresses throughout the proliferative phase of the cycle, to fully restore endometrial thickness. Endometrial repair is unique compared to tissue repair elsewhere in the adult, in that it is rapid, scar-free and occurs around 400 times during each modern woman's reproductive life. The shedding tissue and that undergoing repair is bathed in menstrual fluid, which contains live cells, cellular debris, fragments of extracellular matrix, activated leukocytes and their products, soluble cellular components and extracellular vesicles. Proteomic and other analyses have revealed some detail of these components. Menstrual fluid, along with a number of individual proteins enhances epithelial cell migration to cover the wound. This is shown in endometrial epithelial and keratinocyte cell culture models, in an ex vivo decellularized skin model and in pig wounds in vivo. Thus, the microenvironment provided by menstrual fluid, is likely responsible for the unique rapid and scar-free repair of this remarkable tissue. Insight gained from analysis of this fluid is likely to be of value not only for treating endometrial bleeding problems but also in providing potential new therapies for poorly repairing wounds such as those seen in the aged and in diabetics.

scholarly journals Strain-Programmable Patch for Diabetic Wound Healing

2021 ◽  
Author(s):  
Georgios Theocharidis ◽  
Hyunwoo Yuk ◽  
Heejung Roh ◽  
Liu Wang ◽  
Ikram Mezghani ◽  
...  
Keyword(s):  
Wound Healing ◽  
Chronic Wounds ◽  
Ex Vivo ◽  
Numerical Models ◽  
Diabetic Patients ◽  
Diabetic Wound ◽  
Memory Mechanism ◽  
Culture Models ◽  
Diabetic Wound Healing

Chronic wounds with impaired healing capability such as diabetic foot ulcers (DFU) are devastating complications in diabetic patients, inflicting rapidly growing clinical and economic burdens in aging societies. Despite recent advances in therapeutic approaches, limited benefits of the existing solutions highlight the critical need for novel therapeutic solutions for diabetic wound healing. Here we propose a strain-programmable patch capable of rapid robust adhesion on and programmable mechanical contraction of wet wounded tissues over days to offer a new therapeutic platform for diabetic wounds. The strain-programmable patch, consisting of a dried bioadhesive layer and a pre-stretched elastomer backing, implements a hydration-based shape-memory mechanism to achieve both uniaxial and biaxial contractions and stress remodeling of wet wounds in a programmable manner. We develop theoretical and numerical models to rationally guide the strain-programming and mechanical modulation of wounds. In vivo rodent and ex vivo human skin culture models validate the programmability and efficacy of the proposed platform and identify mechanisms of action for accelerated diabetic wound healing.

scholarly journals 3D culture technologies of cancer stem cells: promising ex vivo tumor models

2020 ◽  
Vol 11 ◽  
pp. 204173142093340 ◽  
Author(s):  
Chengye Zhang ◽  
Zhaoting Yang ◽  
Da-Long Dong ◽  
Tae-Su Jang ◽  
Jonathan C. Knowles ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Ex Vivo ◽  
Three Dimensional ◽  
3D Culture ◽  
Cell Matrix ◽  
Culture Models ◽  
Three Dimensional Culture ◽  
Three Dimensional Models

Cancer stem cells have been shown to be important in tumorigenesis processes, such as tumor growth, metastasis, and recurrence. As such, many three-dimensional models have been developed to establish an ex vivo microenvironment that cancer stem cells experience under in vivo conditions. Cancer stem cells propagating in three-dimensional culture systems show physiologically related signaling pathway profiles, gene expression, cell–matrix and cell–cell interactions, and drug resistance that reflect at least some of the tumor properties seen in vivo. Herein, we discussed the presently available Cancer stem cell three-dimensional culture models that use biomaterials and engineering tools and the biological implications of these models compared to the conventional ones.

scholarly journals Thermodynamics of protein destabilization in live cells

2015 ◽  
Vol 112 (40) ◽  
pp. 12402-12407 ◽  
Author(s):  
Jens Danielsson ◽  
Xin Mu ◽  
Lisa Lang ◽  
Huabing Wang ◽  
Andres Binolfi ◽  
...  
Keyword(s):  
Self Organization ◽  
Live Cells ◽  
Heat Capacity Change ◽  
Bacterial Cells ◽  
Crowding Effects ◽  
Capacity Change ◽  
The Individual ◽  
Cellular Components

Although protein folding and stability have been well explored under simplified conditions in vitro, it is yet unclear how these basic self-organization events are modulated by the crowded interior of live cells. To find out, we use here in-cell NMR to follow at atomic resolution the thermal unfolding of a β-barrel protein inside mammalian and bacterial cells. Challenging the view from in vitro crowding effects, we find that the cells destabilize the protein at 37 °C but with a conspicuous twist: While the melting temperature goes down the cold unfolding moves into the physiological regime, coupled to an augmented heat-capacity change. The effect seems induced by transient, sequence-specific, interactions with the cellular components, acting preferentially on the unfolded ensemble. This points to a model where the in vivo influence on protein behavior is case specific, determined by the individual protein’s interplay with the functionally optimized “interaction landscape” of the cellular interior.

scholarly journals Dynamics of Internalization and Intracellular Interaction of Tau Antibodies and Human Pathological Tau Protein in a Human Neuron-Like Model

2020 ◽  
Vol 11 ◽  
Author(s):  
Dov B. Shamir ◽  
Yan Deng ◽  
Qian Wu ◽  
Swananda Modak ◽  
Erin E. Congdon ◽  
...  
Keyword(s):  
Tau Protein ◽  
Therapeutic Potential ◽  
Ex Vivo ◽  
Human Model ◽  
Paired Helical Filament ◽  
Isotype Control ◽  
Culture Models ◽  
Cellular Compartments ◽  
Cell Culture Models

We and others have shown in various in vivo, ex vivo and cell culture models that several tau antibodies interact with pathological tau within neurons. To further clarify this interaction in a dynamic human model, we differentiated SH-SY5Y cells with retinoic acid and BDNF to create a neuron-like model. Therein, tau antibodies were primarily taken up by receptor-mediated endocytosis, and prevented toxicity of human brain-derived paired helical filament-enriched tau (PHF). Subsequently, we monitored in real-time the interaction of antibodies and PHF within endocytic cellular compartments. Cells were pre-treated with fluorescently-tagged PHF and then incubated with tau antibodies, 4E6, 6B2, or non-specific isotype control IgG1 labeled with a pH sensitive dye. The uptake and binding of the efficacious antibody, 4E6, to PHF occurred mainly within the soma, whereas the ineffective antibody, 6B2, and ineffective control IgG1, were visualized via the processes and showed limited colocalization with PHF within this period. In summary, we have developed a neuron-like model that clarifies the early intracellular dynamics of the interaction of tau antibodies with pathological tau, and identifies features associated with efficacy. Since the model is entirely human, it is suitable to verify the therapeutic potential of humanized antibodies prior to extensive clinical trials.

scholarly journals 3D Organoid Culture From Adult Salivary Gland Tissues as an ex vivo Modeling of Salivary Gland Morphogenesis

2021 ◽  
Vol 9 ◽  
Author(s):  
Donghyun Kim ◽  
Yeo-Jun Yoon ◽  
Dojin Choi ◽  
Jisun Kim ◽  
Jae-Yol Lim
Keyword(s):  
Salivary Gland ◽  
Culture System ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Underlying Mechanism ◽  
Lumen Formation ◽  
Organoid Culture ◽  
Culture Models ◽  
Gland Morphogenesis

Lumen formation of salivary glands has been investigated using in vivo or ex vivo rudiment culture models. In this study, we used a three-dimensional (3D) salivary gland organoid culture system and demonstrated that lumen formation could be recapitulated in mouse SMG organoids. In our organoid culture system, lumen formation was induced by vasoactive intestinal peptide and accelerated by treatment with RA. Furthermore, lumen formation was observed in branching duct-like structure when cultured in combination of fibroblast growth factors (FGF) in the presence of retinoic acid (RA). We suggest RA signaling-mediated regulation of VIPR1 and KRT7 as the underlying mechanism for lumen formation, rather than apoptosis in the organoid culture system. Collectively, our results support a fundamental role for RA in lumen formation and demonstrate the feasibility of 3D organoid culture as a tool for studying salivary gland morphogenesis.

scholarly journals Pharmacological tuning of microRNAs in FAP-derived Extracellular Vesicles by HDAC inhibitors promotes regeneration and reduces fibrosis in dystrophic muscles

2020 ◽  
Author(s):  
M. Sandonà ◽  
S. Consalvi ◽  
L. Tucciarone ◽  
M. De Bardi ◽  
M. Scimeca ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Ex Vivo ◽  
Hdac Inhibitors ◽  
Therapeutic Interventions ◽  
Muscle Stem Cell ◽  
Pathological Conditions ◽  
Regenerative Ability ◽  
Novel Strategy ◽  
Cellular Components

AbstractFunctional interactions between cellular components of the muscle stem cell (MuSC) niche regulate the regenerative ability of skeletal muscles in physiological and pathological conditions; however, the identity of the mediators of these interactions remains largely unknown. We show here that fibro-adipogenic progenitor (FAP)-derived Extracellular Vesicles (EVs) mediate microRNA transfer to MuSCs, and that exposure of dystrophic FAPs to HDAC inhibitors (HDACi) increases the intra-EV levels of a subset of microRNAs (miRs), which cooperatively target biological processes of therapeutic interest, including regeneration, fibrosis and inflammation. In particular, we found that increased levels of miR206 in EVs released from FAPs of muscles from Duchenne dystrophic patients or mice (mdx) exposed to HDACi were associated with enhanced regeneration and inhibition of fibrosis of dystrophic muscles. Consistently, EVs from HDACi-treated dystrophic FAPs could stimulated MuSC activation and expansion ex vivo, and promoted regeneration, while inhibiting fibrosis and inflammation of dystrophic muscles, upon intramuscular transplantation, in vivo. These data reveal a potential for pharmacological modulation of FAP-derived EV’s content as novel strategy for focal therapeutic interventions in Duchenne Muscular Dystrophy (DMD) and possibly other muscular diseases.Brief SummaryExtracellular Vesicles from HDACi-treated dystrophic FAPs promote regeneration, while inhibiting fibrosis and inflammation of dystrophic muscles

scholarly journals In vivo pulse-labeling of isochronic cohorts of cells in the central nervous system using FlashTag

2018 ◽  
Author(s):  
Subashika Govindan ◽  
Polina Oberst ◽  
Denis Jabaudon
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Cell Sorting ◽  
Ex Vivo ◽  
Live Cells ◽  
Fixed Tissue ◽  
System A ◽  
M Phase ◽  
The Central Nervous System

AbstractThis protocol describes a fluorescence birthdating technique to label, track and isolate isochronic cohorts of newborn cells in the central nervous system in vivo. Injection of carboxyfluorescein esters into the cerebral ventricle allows pulse-labeling of M-phase progenitors in touch with the ventricle and their progeny across the central nervous system, a procedure we termed FlashTag. Labeled cells can be imaged ex vivo or in fixed tissue, targeted for electrophysiological experiments, or isolated using Fluorescence-Activated Cell Sorting (FACS) for cell culture or (single-cell) RNA-sequencing. The dye is retained for several weeks, allowing labeled cells to be identified postnatally. This protocol describes the labeling procedure using in utero injection, the isolation of live cells using FACS, as well as the processing of labeled tissue using immunohistochemistry.

Treatment with mTOR Inhibitor, Everolimus (RAD001) Overcomes Resistance to Imatinib In Ph-Leukemia Quiescent Cells.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1579-1579
Author(s):  
Yachiyo Kuwatsuka ◽  
Yosuke Minami ◽  
Miho Minami ◽  
Ryohei Tanizaki ◽  
Hitoshi Kiyoi ◽  
...  
Keyword(s):  
Research Funding ◽  
Mtor Inhibitor ◽  
Ex Vivo ◽  
Nod Mice ◽  
Spleen Cells ◽  
Culture Models ◽  
Nog Mice ◽  
Cord Blood Cells ◽  
Slow Cycling

Abstract Abstract 1579 Recent studies suggest that leukemia stem cells (LSCs) are responsible for relapse of leukemia and eradication of LSCs should be necessary for the cure. In order to examine mechanisms of drug resistance in Ph+ leukemia to imatinib (IM) due to stem cell properties and to seek strategies to overcome the resistance, we've previously established in vivo-murine and ex vivo-culture models using murine hematopoietic pluripotent progenitors transduced with BCR-ABL (Minami, et al., PNAS, 2008). Furthermore, Ph+ leukemia (including T315I-, F311I-mutated CML-BC, or Y253H-mutated Ph-ALL) patient cells were serially xenotransplanted into immunodeficient NOD/SCID/IL2rγnull (NOG) mice. Engrafted bone marrow and spleen cells were almost identical to the original leukemia cells as to phenotypes including karyotypes and distribution of primitive populations. Spleen cells from leukemic NOG mice were treated ex vivo with IM and growth factors, and cell viablility (PI/AnnexinV-staining) was compared between treated and non-treated cells. After treatment with IM, significantly more residual cells were observed in the CD34+/38- population compared to the CD34+/38+ or CD34-/38+ populations. Phosphorylation of BCR-ABL and CrkL was completely inhibited in all populations with IM treatment. Regarding cell cycle states, a higher percentage of quiescent slow-cycling (Hoechst 33342low/Pyronin Ylow) cells was observed in the CD34+/38- population relative to the other populations. Recently, aberrant activation of mTOR signaling has also been reported to be involved in LSCs. Leukemic spleen cells in longer co-culturing with stromal cells were treated with IM and mTOR inhibitor, everolimus (Eve, RAD001). While slow-cycling CD34high+ cells were insensitive to IM, combination treatment with IM and Eve induced significant cell death also in the quiescent population. In vivo-treatment with Eve decreased tumor cells in leukemic NOD mice inoculated with the leukemic NOG cells under the sub-lethally irradiated condition. Treatment with Eve also reduced long-term colony formation of Ph+ leukemia patient samples including Y253H-mutated Ph-ALL with less toxicity to normal cord blood cells. These results imply that treatment with Eve is promising for overcoming the resistance to IM due to quiescent property in Ph+ LSCs. Disclosures: Naoe: Kyowa-Kirin: Research Funding; Novartis: Research Funding; Janssen: Research Funding.

Ex vivo lung culture models to study respiratory inflammation and their relevance for in vivo: An interspecies comparison

2010 ◽  
Vol 196 ◽  
pp. S154
Author(s):  
K. Sewald ◽  
S. Seehase ◽  
S. Switalla ◽  
H.D. Lauenstein ◽  
C. Foerster ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Interspecies Comparison ◽  
Culture Models ◽  
Respiratory Inflammation

scholarly journals Kinomics platform using GBM tissue identifies BTK as being associated with higher patient survival

2021 ◽  
Vol 4 (12) ◽  
pp. e202101054
Author(s):  
Sofian Al Shboul ◽  
Olimpia E Curran ◽  
Javier A Alfaro ◽  
Fiona Lickiss ◽  
Erisa Nita ◽  
...  
Keyword(s):  
Cell Lines ◽  
Drug Targets ◽  
Patient Survival ◽  
Specific Activity ◽  
Ex Vivo ◽  
Culture Models ◽  
Novel Drug ◽  
Btk Inhibitors ◽  
Novel Drug Targets

Better understanding of GBM signalling networks in-vivo would help develop more physiologically relevant ex vivo models to support therapeutic discovery. A “functional proteomics” screen was undertaken to measure the specific activity of a set of protein kinases in a two-step cell-free biochemical assay to define dominant kinase activities to identify potentially novel drug targets that may have been overlooked in studies interrogating GBM-derived cell lines. A dominant kinase activity derived from the tumour tissue, but not patient-derived GBM stem-like cell lines, was Bruton tyrosine kinase (BTK). We demonstrate that BTK is expressed in more than one cell type within GBM tissue; SOX2-positive cells, CD163-positive cells, CD68-positive cells, and an unidentified cell population which is SOX2-negative CD163-negative and/or CD68-negative. The data provide a strategy to better mimic GBM tissue ex vivo by reconstituting more physiologically heterogeneous cell co-culture models including BTK-positive/negative cancer and immune cells. These data also have implications for the design and/or interpretation of emerging clinical trials using BTK inhibitors because BTK expression within GBM tissue was linked to longer patient survival.

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