1) Physical confinement prolongs the duration of mitosis in cells, with more confinement leading to longer mitosis. Confinement increases the frequency of mitotic arrest and cell death.
2) Microfluidic devices with variable channel widths were used to study mitosis in confined environments that mimic physiological spaces. Cancer cells and fibroblasts underwent prolonged and irregular mitosis in narrower channels.
3) The mechanisms of mitotic delay in confinement may involve insufficient space for chromosome alignment or incomplete attachment of the spindle to chromosomes. Future work aims to better understand these mechanisms and examine the effects of confinement on non-cancerous cells.
Development of cancer therapeutics is often carried out in 2D cultures prior to testing on animal model. In comparison to 2D cultures, discuss the potential of using 3D in vitro models for drug efficiency testing.
Kim Solez A renaissance in renal pathology brought about by regenerative medi...Kim Solez ,
Dr. Kim Solez presents A renaissance in renal pathology, nephrology and transplantation brought about by regenerative medicine: How to jump start the process.
Development of cancer therapeutics is often carried out in 2D cultures prior to testing on animal model. In comparison to 2D cultures, discuss the potential of using 3D in vitro models for drug efficiency testing.
Kim Solez A renaissance in renal pathology brought about by regenerative medi...Kim Solez ,
Dr. Kim Solez presents A renaissance in renal pathology, nephrology and transplantation brought about by regenerative medicine: How to jump start the process.
Cancer Stem Cells and the Unicellular Life Cycle of Cancer_Crimson PublishersCrimsonpublishersCancer
All eukaryotes, from protists to mammalians, preserve a unicellular life cycle inherited from the common ancestor that can be reactivated under unfavorable living conditions. The cell-of-origin of cancer escapes its death by forming a protected polyploid cyst-like structure (CLS), that starts the unicellular life cycle of cancer. The reversal to unicellularity occurs through genomic and epigenetic alterations that activate the MUT switch of early Metazoans and not through mutations. The microcell progeny of CLSs spread into tissues and organs and form the CSC pool of aCLS cancers. Depending on the environment, the CSC pool differentiates a reproductive cell subline, which forms new aCLSs by cyclic encystment and asymmetric cell division, or a somatic subline, which proliferates strongly by symmetric cell division without cyst differentiation.
Kim Solez Renal transplant pathology and future perspectivesKim Solez ,
Dr. Kim Solez presents "Renal transplant pathology and future perspectives" as a TTS webinar on Dec. 8 at noon EST . Includes discussion of the new discipline of tissue engineering pathology. https://www.tts.org/education/advanced-renal-transplantation
Circulating microRNAs predict initial of NHL in a novel in vivo model: impact...Laura Berry
Presented at the 3rd qPCR and Digital PCR Congress: USA. To find out more, visit:
www.global-engage.com
Afshin Beheshti is an Assistant Professor at the Tufts University School of Medicine. In this presentation, Afshin discusses his investigation into age-dependent circulating microRNA signatures that may influence non-Hodgkin lymphoma.
Liquid biopsies- Learn about the newer ways of diagnosing malignancyBabu Appat
Liquid biopsies are innovative methods of detecting serious conditions at its very onset. It's now in its developmental stage. Some of the liquid biopsies are approved by the FDA of United States and are spreading across the world. Let's hope that this opens the field of easier, earlier and more economic detection of diseases.
Cancer Stem Cells and the Unicellular Life Cycle of Cancer_Crimson PublishersCrimsonpublishersCancer
All eukaryotes, from protists to mammalians, preserve a unicellular life cycle inherited from the common ancestor that can be reactivated under unfavorable living conditions. The cell-of-origin of cancer escapes its death by forming a protected polyploid cyst-like structure (CLS), that starts the unicellular life cycle of cancer. The reversal to unicellularity occurs through genomic and epigenetic alterations that activate the MUT switch of early Metazoans and not through mutations. The microcell progeny of CLSs spread into tissues and organs and form the CSC pool of aCLS cancers. Depending on the environment, the CSC pool differentiates a reproductive cell subline, which forms new aCLSs by cyclic encystment and asymmetric cell division, or a somatic subline, which proliferates strongly by symmetric cell division without cyst differentiation.
Kim Solez Renal transplant pathology and future perspectivesKim Solez ,
Dr. Kim Solez presents "Renal transplant pathology and future perspectives" as a TTS webinar on Dec. 8 at noon EST . Includes discussion of the new discipline of tissue engineering pathology. https://www.tts.org/education/advanced-renal-transplantation
Circulating microRNAs predict initial of NHL in a novel in vivo model: impact...Laura Berry
Presented at the 3rd qPCR and Digital PCR Congress: USA. To find out more, visit:
www.global-engage.com
Afshin Beheshti is an Assistant Professor at the Tufts University School of Medicine. In this presentation, Afshin discusses his investigation into age-dependent circulating microRNA signatures that may influence non-Hodgkin lymphoma.
Liquid biopsies- Learn about the newer ways of diagnosing malignancyBabu Appat
Liquid biopsies are innovative methods of detecting serious conditions at its very onset. It's now in its developmental stage. Some of the liquid biopsies are approved by the FDA of United States and are spreading across the world. Let's hope that this opens the field of easier, earlier and more economic detection of diseases.
The Acoustic Technology for Ctcs Isolation in Blood: Low-Cost Devices_Crimson...CrimsonpublishersCancer
Blood samples can be used as a liquid biopsy in cancer diagnosis and chemotherapy monitoring. This label- free method offers benefits over traditional tissue invasive biopsy. It is possible to separate rare cells from blood samples by Ultrasounds on the basis of their physical properties in a biocompatible manner. A successful separation of cultured cancer cells from WBCs with acoustic-based methods is being demonstrated during the last years through different technological approaches. The concept of plate acoustic waves (PAW) applied to acoustophoresis was recently introduced to perform acoustic flow-through separation of rare cells in blood samples. It lies in the geometrical chip design, different to other micro separators (BAW and SAW). This new strategy allows soft materials of extremely reduced volume and low-cost fabrication and opens a door to printing manufacturing processes.
[論文紹介] 古代の伝染性がん系統の体細胞進化と世界的拡大 (Somatic evolution and global expansion of an a...Shohei Nagata
論文紹介 (2019)
Somatic evolution and global expansion of an ancient
transmissible cancer lineage
古代の伝染性がん系統の体細胞進化と世界的拡大
※なぜか文字が表示されない場合は全画面表示や保存をすると表示されるようです。
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Visualizing Human Stem Cell Dynamics by Multicolor, Multiday High-Content Mic...InsideScientific
Visualizing the complex spatiotemporal dynamics of human stem cells as they proliferate and make cell fate decisions is key to improve our understanding of how to robustly engineer differentiated tissues for therapeutic applications.
In this webinar, Dr. Rafael Carazo Salas describes multicolor, multiday high-content microscopy pipelines that his group has recently developed to visualize the dynamical cell fate changes of human Pluripotent Stem Cells (hPSCs). In particular, he reviews the integrated experimental and computational approaches that his group has established, including novel “live” reporters of cell fate and multi-reporter hPSC lines generated by CRISPR/Cas9 allowing multiplexed monitoring of cell proliferation and fate dynamics, and exemplify the biological discoveries they are enabling.
Key Topics Include:
- Visualizing how human Pluripotent Stem Cells (hPSCs) proliferate and undergo early differentiation in vitro, by high content microscopy
- Learning about experimental and computational pipelines that enable cell fate monitoring at the collective and single-cell level
- Learning about novel “live” reporters of hPSC cell fate
Genes and Tissue Culture Technology Assignment (G6)Rohini Krishnan
The culture of cells in two dimensions does not reproduce the histological characteristics of a tissue for informative or useful study. Growing cells as three-dimensional (3D) models more analogous to their existence in vivo may be more clinically relevant.
REVIEWCancer stem cells a new framework for the designo.docxjoellemurphey
REVIEW
Cancer stem cells: a new framework for the design
of tumor therapies
Boyan K. Garvalov & Till Acker
Received: 14 July 2010 /Revised: 27 August 2010 /Accepted: 16 September 2010
# Springer-Verlag 2010
Abstract Modern tumor therapy has achieved considerable
progress, but many tumors remain refractory to treatment or
relapse following initial remission. Recent evidence points
to one possible reason for this limited therapeutic efficiency:
that the design of anticancer agents so far may not have been
aimed at the right target. While conventional tumor therapies
have targeted the main mass of tumor cells, there is now
compelling evidence that tumor initiation and progression are
driven by a subpopulation of tumor cells that possess stem cell
properties and are resistant to traditional cancer treatments—
the cancer stem cells (CSCs). CSCs have been identified in
most types of cancer and can be separated from the rest of the
tumor cells using appropriate markers. CSCs are regulated by
molecular mechanisms and specific, perivascular, and hypox-
ic microenvironments, which largely overlap with those
controlling stem cells from normal tissues. Our improved
understanding of CSC biology has already provided a number
of novel targets and drug discovery platforms for the design of
specific therapies that aim to eradicate the CSC subpopula-
tion. Therapeutic approaches can be targeted either at
eliminating the CSCs themselves or at disrupting the niches
in which CSCs reside. Moreover, the importance of CSCs for
tumor growth, resistance, and progression implies that clinical
trials and preclinical studies of anticancer therapies should
include as a key element an assessment of the abundance and
persistence of CSCs. Thus, CSC research holds great promise
for providing important new impetus to the fields of tumor
biology and clinical oncology.
Keywords Cancer stem cell . Hypoxia .
Microenvironment . Angiogenesis . Antitumor therapy.
Metastasis
The hierarchy model and cancer stem cells (CSCs)
The classical view of tumor formation is based on the
“stochastic” or “clonal evolution” model [1, 2]. It perceives
the tumor as a mass of hyperproliferative cells with similar
potential for driving tumor growth. Tumor heterogeneity
and progression are seen as the result of variations in the
tumor microenvironment and genetic mutations in individ-
ual cells, followed by selection of those that are best
adapted to support the further growth of the tumor (Fig. 1a).
An alternative concept that has been gaining increasing
experimental support is the “hierarchy” or “cancer stem
cell” model [3]. This model posits that tumors are generated
and maintained in a manner similar to the physiological
stem cell system operating in normal tissues, i.e., by cells
with stem cell-like properties, which self-renew and
differentiate into the distinct cellular subtypes of the tumor
(Fig. 1b). The key novel features of this model are that only
a limited population of tumor cell ...
Genome folding by loop extrusion and compartmentalization Leonid Mirny
2018-03-27 Leonid Mirny (MIT) and Nezar Abdennur, Sameer Abraham, Ed Banigan, Hugo Brandao, Martin Falk, Geoff Fudenberg (UCSF), Anton Goloborodko, Max Imakaev, Carolyn Lu, Johannes Nuebler, Aafke van den Berg; talk at the Keystone Symposium
Mammalian MSC from Selected Species: Features and Applications
Christiane Uder, Sandra Br€uckner, Sandra Winkler, Hans-Michael Tautenhahn,†‡ Bruno Christ†*
Mesenchymal stromal/stem cells (MSC) are promising candidates for cellular therapy of different diseases in humans and in animals. Following the guidelines of the International Society for Cell Therapy, human MSC may be identified by expression of a specific panel of cell surface markers (CD1051, CD731, CD901, CD34-, CD14-, or CD11b-, CD79- or CD19-, HLA-DR-). In addition, multiple differentiation potential into at least the osteogenic, adipogenic, and chondrogenic lineage is a main criterion for MSC definition. Human MSC and MSC of a variety of mammals isolated from different tissues meet these criteria. In addition to the abovementioned, they express many more cell surface markers. Yet, these are not uniquely expressed by MSC. The gross phenotypic appearance like marker expression and differentiation potential is similar albeit not identical for MSC from different tissues and species. Similarly, MSC may feature different biological characteristics depending on the tissue source and the isolation and culture procedures. Their versatile biological qualities comprising immunomodulatory, anti-inflammatory, and proregenerative capacities rely largely on the migratory and secretory capabilities of MSC. They are attracted to sites of tissue lesion and secrete factors to promote self-repair of the injured tissue. This is a big perspective for clinical MSC applications in both veterinary and human medicine. Phase I/II clinical trials have been initiated to assess safety and feasibility of MSC therapies in acute and chronic disease settings. Yet, since the mode of MSC action in a specific disease environment is still unknown at large, it is mandatory to unravel the response of MSC from a given source onto a specific disease environment in suitable animal models prior to clinical applications.
Mammalian MSC from Selected Species: Features and Applications Christiane Ude...
Cassidy Wang Poster Final
1. Physically Confined Microenvironments
Impair Mitotic Progression
Cassidy Wang, Xiaohu Wan, Konstantinos Konstantopoulos
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA
0
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30 60 90 120 150 180 210 240 270 300 330 360 390 420 450 480 510 540
Frequency (%)
Duration (minutes)
HT1080 Mitosis Duration Distribution
Unconfined
10 μm channel
6 μm channel
3 μm channel
Introduction
Conclusions
Elevated Incidence of Arrest, Delay, and
Death in Mitosis
Mitosis in Confinement
Future Directions
Acknowledgments
I extend my thanks to Konstantinos Konstantopoulos and Xiaohu Wan for
their support and mentorship throughout this project. My appreciation goes
to Chris Yankaskas for his patience and guidance. I am grateful to all the
members of the Kostas Lab for making my time in the lab both enlightening
and immensely enjoyable. Lastly, I would like to thank INBT for making this
experience possible.
References
Critical to understanding confined cell dynamics is the characterization of
mitosis. Mitosis is required for tissue development and has been shown to
differ between confined and unconfined environments. In vivo studies have
also suggested that intravascular tumor cell division plays a critical role in
cancer metastasis.1 The importance of physically confined mitosis in tissue
growth and cancer progression serves as the motivation for this research
project.
Platform for Confinement Studies
• PDMS microchannel devices
• ECM protein (collagen-I) applied to device
• Variable channel widths
• Enables easy imaging of microchannels
Confinement Prolongs Cell Division
Relationship Between Degree of
Confinement And Mitosis Duration
• Physical confinement prolongs mitosis
• Degree of confinement is directly related to duration of mitosis
• Confinement increases frequency of mitotic arrest and apoptosis
• Identify the mechanism of mitotic delay in confinement
• Conduct confocal imaging of cells with chromatin and tubulin labeling
• Determine number of chromosomes during confined mitosis
• Examine relationship between migratory ability and duration of mitosis
• Study confined mitosis in non-cancerous cell lines
• Forms bipolar spindle similar to that seen in unconfined mitosis
• More elongated morphology
1. Xi, W., Schmidt, C. K., Sanchez, S., Gracias, D. H., Carazo-Salas, R. E., Butler, R., ...
Schmidt, O. G. (2016). Molecular Insights into Division of Single Human Cancer Cells in
On-Chip Transparent Microtubes. ACS Nano, 10(6), 5835-5846.
2. Hung, W.-C., Chen, S.-H., Paul, C. D., Stroka, K. M., Lo, Y.-C., Yang, J. T., &
Konstantopoulos, K. (2013). Distinct signaling mechanisms regulate migration in
unconfined versus confined spaces. The Journal of Cell Biology, 202(5), 807–824.
Experimental Design
Classical cell biology studies are conducted on flat surfaces that pose little to
no obstacle to cell proliferation. While the chemical environments of these
“two-dimensional” surfaces can be made to mimic physiological conditions,
they do not accurately replicate the physical spaces in which cells grow and
spread. By using microfluidic devices, cell proliferation and migration can be
studied in varying levels of confinement that more accurately represent
physiological spaces. Examination of cell dynamics in 3D confined
microenvironments has far-reaching implications in regenerative medicine,
drug efficacy and toxicity assays, and cancer treatment.
Figure 1. Cancer
cells in confinement.
Xi et al. (2016)1
0
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30 60 90 120 150 180 210 240 270 300 330 360 390 420
Frequency (%)
Duration (minutes)
MDA-MB-231 Mitosis Duration Distribution
Unconfined
3 μm channel
Mitosis in unconfined space. HeLa cells stained with SiR-Tubulin (green) and
mCherry H2B (red). (Daniel Gerlich and Claudia Blaukopf, Institute of Molecular
Biotechnology, Vienna)
• Fluid pressure generates flow
• Cells adhere to channel entrances
• Cells migrate through channels
• Upper wells may be used to establish chemotactic gradient
Changes in cell morphology with
varying degrees of confinement.
Hung et al. (2013)2
Cells
Pressure-driven
flow
• 3, 6, 10 μm width
• 1 mm length
• 10 μm height
• 3 μm width
• 200 μm length
• 10 μm height
Fluorescence images
(tubulin)
Phase images
65
158
0
50
100
150
200
250
300
Unconfined 3 μm channel
Duration (minutes)
Average Duration of MDA-MB-231 Mitosis Under Physical Confinement
N=74 N=25
6 μm channel
47
77
130
348
0
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300
400
500
600
Unconfined 10 μm channel 6 μm channel 3 μm channel
Duration (minutes)
Average Duration of HT1080 Mitosis Under Physical Confinement
N=53 N=62 N=37 N=11
Hypothesis
• Insufficient space for chromosome alignment
• Incomplete attachment of spindle to chromosomes
0
11
35
97
0
20
40
60
80
100
Unconfined 10 μm channel 6 μm channel 3 μm channel
Percentage of Mitotic Events (%)
Relative Frequencies of Mitotic Delays/Deaths
Arrest/delay
threshold:
3 hours
N=53 N=66 N=46 N=30
Migration
Device designs used in study