Combined influence of biophysical and biochemical cues on maintenance and proliferation of hematopoietic stem cells

Gvaramia, David; Müller, Eike; Müller, Karl H.; Atallah, Passant; Tsurkan, Mikhail V.; Freudenberg, Uwe; Bornhäuser, Martin; Werner, Carsten

doi:10.1016/j.biomaterials.2017.05.023

Cited by 49 publications

(45 citation statements)

References 50 publications

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“…In the current study, a decrease in proliferation under spatial confinement was observed for all cell lines tested over one to three days in the confiner. Similar results were found by [49], who reported a higher frequency of quiescent cells upon confinement within a stiff 3D matrix. These results also corroborate those obtained using PDMS set-ups [50,51] and atomic force microscopy (AFM) [52], where it was reported that confinement delays mitotic progression.…”

Section: Discussionsupporting

confidence: 89%

“…We anticipate that this device could be a valuable tool for the fundamental understanding of the effect of cell confinement on various hallmarks of cancer progression and resistance, and in particular to decipher the role of nuclear mechanosensitivity. Links between cell resistance to treatment and mechanical stimuli have been highlighted recently [49,67,68]. Such mechanical cues could hence lead to new therapeutic approaches [69] that could be tested using such systems.…”

Section: Acknowledgmentsmentioning

confidence: 99%

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Development of a soft cell confiner to decipher the impact of mechanical stimuli on cells

Prunet

Lefort

Delanoë-Ayari

et al. 2020

Preprint

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Emerging evidence suggests the importance of mechanical stimuli in normal and pathological situations for the control of many critical cellular functions. While the effect of matrix stiffness has been and is still extensively studied, few studies have focused on the role of mechanical stresses. The main limitation of such analyses is the lack of standard in vitro assays enabling extended mechanical stimulation compatible with dynamic biological and biophysical cell characterization. We have developed an agarose-based microsystem, the soft cell confiner, which enables the precise control of confinement for single or mixed cell populations. The rigidity of the confiner matches physiological conditions and enables passive medium renewal. It is compatible with time-lapse microscopy, in situ immunostaining, and standard molecular analyses, and can be used with both adherent and nonadherent cell lines. Cell proliferation of various cell lines (hematopoietic cells, MCF10A epithelial breast cells and HS27A stromal cells) was followed for several days up to confluence using video-microscopy and further documented by Western blot and immunostaining. Interestingly, even though the nuclear projected area was much larger upon confinement, with many highly deformed nuclei (non-circular shape), cell viability, assessed by live and dead cell staining, was unaffected for up to 8 days in the confiner. However, there was a decrease in cell proliferation upon confinement for all tested cell lines. The soft cell confiner is thus a valuable tool to decipher the effect of long-term confinement and deformation on the biology of cell populations. This tool will be instrumental in deciphering the impact of nuclear and cytoskeletal mechanosensitivity in normal and pathological conditions involving highly confined situations, such as those reported upon aging with fibrosis or during cancer. Highlights-We have developed a flexible hydrogel-based microsystem that precisely confines cells for extended periods of time with controlled nutrient levels.-The soft confiner system was validated for both adherent and non-adherent cells and allows the simultaneous in situ follow-up or ex situ analysis of multiple subpopulations.-A unique tool to analyze the role of long-term effect of mechanical confinement in normal and pathological conditions. Graphical abstract3/27

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Section: Discussionsupporting

confidence: 89%

Section: Acknowledgmentsmentioning

confidence: 99%

Development of a soft cell confiner to decipher the impact of mechanical stimuli on cells

Prunet

Lefort

Delanoë-Ayari

et al. 2020

Preprint

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“…Moreover, an osmotic compression, stress of very different origin and sensing which could arise in vivo due to accumulation of oncotic pressure [27], also leads to similar modulation of chemotherapeutic. There are now evidences that quiescence is one major mechanism leading to drug resistance [28] and that mechanics can turn cells towards quiescence [8,29]. However, to our best knowledge, no quantitative models linking mechanics to drug resistance, through a direct modulation of proliferation, have been proposed so far.…”

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confidence: 99%

Mechanical-control of cell proliferation increases resistance to chemotherapeutic agents

Rizzuti¹,

Mascheroni²,

Arcucci³

et al. 2020

Preprint

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While many cellular mechanisms leading to chemotherapeutic resistance have been identified, there is an increasing realization that tumor-stroma interactions also play an important role. In particular, mechanical alterations are inherent to solid cancer progression and profoundly impact cell physiology. Here, we explore the impact of compressive stress on the efficacy of chemotherapeutics in pancreatic cancer spheroids. We find that increased compressive stress leads to decreased drug efficacy. Theoretical modeling and experiments suggest that mechanical stress leads to decreased cell proliferation which in turn reduces the efficacy of chemotherapeutics that target proliferating cells. Our work highlights a mechanical-form of drug resistance, and suggests new strategies for therapy.Mechanical alterations of solid tumors are a hallmark of cancer progression. Among the many occurring modifications, the most representative forms of mechanical alterations in tumors are changes in extracellular matrix rigidity[1] and build up of compressive stress [2]. Compressive stress accumulation can be found in many cancers such as glioblastoma multiform [3] or pancreatic ductal adenocarcinoma (PDAC) [2].PDAC is one of the deadliest cancers, with extremely poor prognosis when diagnosed and no efficient treatment available besides surgery. PDAC development is characterized by excessive deposition of extracellular material during which strong modifications of the mechanical environment arises. In particular, the deposition of negatively-charged hyaluronic acid leads to electroswelling of extracellular matrix and subsequent compressive stress experienced by tumor cells [4]. The local growth of cancer cells in an elastic environment also leads to build-up of compressive stress through a process known as growth-induced pressure [5,6]. PDAC tumors are extremely compressed, within the kPa range [7].In vitro, compressive stress can alter cell physiology in multiple ways, from proliferation [8,9] to migration [10,11]. In vivo, it has recently been shown that compressive stress built-up in PDAC tumors can exceed blood pressure and participate in blood vessel collapse [12]. Most of large vessels (diameter above 10µm) are clamped, leading to poor perfusion. Vessel collapse is associated with drug resistance: classical first-line chemotherapeutics such as gemcitabine are thought to be unable to reach the tumor, which decreases or even prevents the effect of the drug. Intravenous injection of a pegylated-form of hyaluronidase, an enzyme digesting hyaluronic acid, renormalizes blood vessels and, in combination with gemcitabine, increases chemotherapeutic efficacy [12].The proposed mechanism overcoming this form of resistance is better tumor perfusion through decrease in compressive stress. However, it remains unclear how the combination of hyaluronidase and gemcitabine really works. Indeed, if the vessels are so collapsed that gemcitabine does not penetrate the tumor, hyaluronidase should not have a better chance to reach the tumor. Another pot...

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“…A number of recent studies have focused on mimicking the BM niche characteristics in vitro by creating 3D culture systems that incorporate, along with growth factors and cytokines, different biomaterials to provide a range of biophysical cues in an attempt to induce symmetric cell divisions of HSCs in vitro [ 26 , 27 , 28 , 29 , 30 , 31 , 32 ]. Biomaterial elasticity is currently recognized to affect the fate of HSPCs [ 33 , 34 , 35 , 36 ]. In 2010, Holst et al reported that the proliferation of repopulating HSCs was enhanced when cells were cultured on top of soft tropoelastin-coated plates compared to cells cultured on top of control plates without tropoelastin [ 33 ].…”

Section: Introductionmentioning

confidence: 99%

Towards Mimicking the Fetal Liver Niche: The Influence of Elasticity and Oxygen Tension on Hematopoietic Stem/Progenitor Cells Cultured in 3D Fibrin Hydrogels

Abrego

Zaunz

Toprakhisar

et al. 2020

IJMS

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Hematopoietic stem/progenitor cells (HSPCs) are responsible for the generation of blood cells throughout life. It is believed that, in addition to soluble cytokines and niche cells, biophysical cues like elasticity and oxygen tension are responsible for the orchestration of stem cell fate. Although several studies have examined the effects of bone marrow (BM) niche elasticity on HSPC behavior, no study has yet investigated the effects of the elasticity of other niche sites like the fetal liver (FL), where HSPCs expand more extensively. In this study, we evaluated the effect of matrix stiffness values similar to those of the FL on BM-derived HSPC expansion. We first characterized the elastic modulus of murine FL tissue at embryonic day E14.5. Fibrin hydrogels with similar stiffness values as the FL (soft hydrogels) were compared with stiffer fibrin hydrogels (hard hydrogels) and with suspension culture. We evaluated the expansion of total nucleated cells (TNCs), Lin−/cKit+ cells, HSPCs (Lin−/Sca+/cKit+ (LSK) cells), and hematopoietic stem cells (HSCs: LSK- Signaling Lymphocyte Activated Molecule (LSK-SLAM) cells) when cultured in 5% O2 (hypoxia) or in normoxia. After 10 days, there was a significant expansion of TNCs and LSK cells in all culture conditions at both levels of oxygen tension. LSK cells expanded more in suspension culture than in both fibrin hydrogels, whereas TNCs expanded more in suspension culture and in soft hydrogels than in hard hydrogels, particularly in normoxia. The number of LSK-SLAM cells was maintained in suspension culture and in the soft hydrogels but not in the hard hydrogels. Our results indicate that both suspension culture and fibrin hydrogels allow for the expansion of HSPCs and more differentiated progeny whereas stiff environments may compromise LSK-SLAM cell expansion. This suggests that further research using softer hydrogels with stiffness values closer to the FL niche is warranted.

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Combined influence of biophysical and biochemical cues on maintenance and proliferation of hematopoietic stem cells

Cited by 49 publications

References 50 publications

Development of a soft cell confiner to decipher the impact of mechanical stimuli on cells

Development of a soft cell confiner to decipher the impact of mechanical stimuli on cells

Mechanical-control of cell proliferation increases resistance to chemotherapeutic agents

Towards Mimicking the Fetal Liver Niche: The Influence of Elasticity and Oxygen Tension on Hematopoietic Stem/Progenitor Cells Cultured in 3D Fibrin Hydrogels

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