Abstract 120: Identification Of A Novel Adult Smooth Muscle-like Stem/progenitor Cell That Facilitates Formation Of Tissue Engineered Vascular Tissue For Use As Vascular Grafts In Vivo

The aim of this study was to characterise a recently identified adult smooth muscle stem-like/progenitor cell (SMSPC) and exploit the ability to differentiate contractile smooth muscle cells (SMC) from SMSPCs facilitating tissue engineering of small to medium arteries, an approach which is hampered by inadequate sources of, and scale up methodologies for autologous SMCs. We initially identified a novel SMSPC in the bone marrow (BM) of rats that retained classic stem cell-like characteristics (clonogenicity, unlimited self-renewal, high telomerase activity, expression of stem cell markers) whilst expressing low levels of SMC-like transcripts. Spontaneous differentiation favoured a SMC phenotype where ~20-fold increases in calponin and SM-MHC protein expression were observed 10 days post-embryoid body formation, an effect augmented by TGFβ (P < 0.0001). Differentiation of SMSPCs from an undifferentiated state towards contractile SMCs was Myocd-dependent. Overexpression of Myocd in SMSPCs increased their percentage contraction in response to AngII (P < 0.05) and KCL (P < 0.05) and Ca2+ signalling to levels indistinguishable from rat SMCs. Knockdown of SMC-specific repressors resulted in Myocd-dependent SMC differentiation of SMSPCs. Taking advantage of the fact that SMSPCs possessed unlimited self-renewal capacity ex vivo and could be successfully reprogrammed into functional SMCs, we determined the vascular tissue engineering potential of SMSPCs and showed these cells integrate as viable tissue engineered vascular grafts in vivo. Specifically, after 1 month implantation, a Myocd-GFP+/Isl1+ smooth muscle layer was observed in the grafts along with a vWF+ luminal endothelial layer and multi-layered collagen and elastin fibre formation throughout the graft wall. We have recently isolated cells with similar SMSPC phenotype and differentiation potential from human peripheral blood. This study demonstrates that adult SMSPCs derived from rat BM and human blood can be reprogrammed to efficiently generate large quantities of mature contractile SMCs, displaying great utility as a cellular backbone for tissue engineered vascular grafts, making them an attractive source for vascular cell therapy and surgical applications.

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Surface Modification by Nanobiomaterials for Vascular Tissue Engineering Applications

Current Medicinal Chemistry ◽

10.2174/0929867325666180914104633 ◽

2020 ◽

Vol 27 (10) ◽

pp. 1634-1646 ◽

Cited By ~ 1

Author(s):

Huey-Shan Hung ◽

Shan-hui Hsu

Keyword(s):

Tissue Engineering ◽

Vascular Tissue ◽

Thrombus Formation ◽

Vascular Grafts ◽

Vascular Tissue Engineering ◽

Great Success ◽

Natural Polymers ◽

Vascular Biomaterials

Treatment of cardiovascular disease has achieved great success using artificial implants, particularly synthetic-polymer made grafts. However, thrombus formation and restenosis are the current clinical problems need to be conquered. New biomaterials, modifying the surface of synthetic vascular grafts, have been created to improve long-term patency for the better hemocompatibility. The vascular biomaterials can be fabricated from synthetic or natural polymers for vascular tissue engineering. Stem cells can be seeded by different techniques into tissue-engineered vascular grafts in vitro and implanted in vivo to repair the vascular tissues. To overcome the thrombogenesis and promote the endothelialization effect, vascular biomaterials employing nanotopography are more bio-mimic to the native tissue made and have been engineered by various approaches such as prepared as a simple surface coating on the vascular biomaterials. It has now become an important and interesting field to find novel approaches to better endothelization of vascular biomaterials. In this article, we focus to review the techniques with better potential improving endothelization and summarize for vascular biomaterial application. This review article will enable the development of biomaterials with a high degree of originality, innovative research on novel techniques for surface fabrication for vascular biomaterials application.

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Resveratrol Suppresses Epithelial-Mesenchymal Transition in GBM by Regulating Smad-Dependent Signaling

BioMed Research International ◽

10.1155/2019/1321973 ◽

2019 ◽

Vol 2019 ◽

pp. 1-14 ◽

Cited By ~ 10

Author(s):

Yang Song ◽

Yong Chen ◽

Yunqian Li ◽

Xiaoyan Lyu ◽

Jiayue Cui ◽

...

Keyword(s):

Stem Cell ◽

Epithelial Mesenchymal Transition ◽

Inhibitory Effect ◽

Stem Cell Markers ◽

Invasive Ability ◽

Self Renewal ◽

Invasive Phenotype ◽

Mesenchymal Transition ◽

Cancer Stem Cell Markers

Glioblastoma (GBM) is the most common and malignant intracranial tumor in adults. Despite continuous improvements in diagnosis and therapeutic method, the prognosis is still far away from expectations. The invasive phenotype of GBM is the main reason for the poor prognosis. Epithelial-mesenchymal transition (EMT) is recognized as a participator in this invasive phenotype. Resveratrol, a natural plant-derived compound, is reported to be able to regulate EMT. In the present study, we used TGF-β1 to induce EMT and aimed to evaluate the effect of resveratrol on EMT and to explore the underline mechanism in GBM. Western blotting was used to detect the expression of EMT-related markers, stemness markers, and Smad-dependent signaling. Wound healing assay and transwell invasion assay were performed to evaluate the migratory and invasive ability of GBM cells. Gliosphere formation assay was used to investigate the effect of resveratrol on the ability of self-renewal. Xenograft experiment was conducted to examine the effect of resveratrol on EMT and Smad-dependent signalingin vivo. Our data validated that resveratrol suppressed EMT and EMT-associated migratory and invasive ability via Smad-dependent signaling in GBM cells. We also confirmed that resveratrol obviously inhibited EMT-induced self-renewal ability of glioma stem cells (GSCs) and inhibited EMT-induced cancer stem cell markers Bmi1 and Sox2, suggesting that resveratrol is able to suppress EMT-generated stem cell-like properties in GBM cells. Furthermore, we also showed the inhibitory effect of resveratrol on EMT in xenograft experimentsin vivo. Overall, our study reveals that resveratrol suppresses EMT and EMT-generated stem cell-like properties in GBM by regulating Smad-dependent signaling and provides experimental evidence of resveratrol for GBM treatment.

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Spatial Distributions, Characteristics, and Applications of Craniofacial Stem Cells

Stem Cells International ◽

10.1155/2020/8868593 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Geru Zhang ◽

Qiwen Li ◽

Quan Yuan ◽

Shiwen Zhang

Keyword(s):

Stem Cells ◽

Spatial Distribution ◽

Stem Cell ◽

Cranial Sutures ◽

Stem Cell Markers ◽

Spatial Distributions ◽

Self Renewal ◽

Stem Cell Niches ◽

Multidirectional Differentiation

Stem cells play an irreplaceable role in the development, homeostasis, and regeneration of the craniofacial bone. Multiple populations of tissue-resident craniofacial skeletal stem cells have been identified in different stem cell niches, including the cranial periosteum, jawbone marrow, temporomandibular joint, cranial sutures, and periodontium. These cells exhibit self-renewal and multidirectional differentiation abilities. Here, we summarized the properties of craniofacial skeletal stem cells, based on their spatial distribution. Specifically, we focused on the in vivo genetic fate mapping of stem cells, by exploring specific stem cell markers and observing their lineage commitment in both the homeostatic and regenerative states. Finally, we discussed their application in regenerative medicine.

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ID: 22: DIFFERENTIAL ACTIONS OF ESTROGEN RECEPTOR α AND β VIA NON-GENOMIC SIGNALING IN HUMAN PROSTATE STEM-PROGENITOR CELLS

Journal of Investigative Medicine ◽

10.1136/jim-2016-000120.46 ◽

2016 ◽

Vol 64 (4) ◽

pp. 933-933

Author(s):

S Majumdar ◽

JC Rinaldi ◽

T Gauntner ◽

L Xie ◽

W Hu ◽

...

Keyword(s):

Cell Proliferation ◽

Stem Cell ◽

Cell Line ◽

Progenitor Cells ◽

Progenitor Cell ◽

Mapk Pathway ◽

Human Prostate ◽

Stem Cell Markers ◽

Self Renewal ◽

Progenitor Cell Proliferation

Genomic signaling via estrogen receptors (ER) has been widely studied and implicated as the main ER signaling pathway in prostate development and carcinogenesis. Non-genomic ER signaling has also been reported in prostate epithelium although down-stream cascades have not been clarified. Our lab has recently identified ERs in human prostate epithelial stem/progenitor cells and shown that that 17β-estradiol (E2) can stimulate stem cell symmetric self-renewal and progenitor cell proliferation. In this study we interrogate non-genomic membrane initiated ER signaling in this prostate stem/progenitor cell population. Human prostate stem-progenitor cells were enriched from primary prostate epithelial cell cultures (PrEC) of young, disease-free donors using a 3D prostasphere (PS) model as previously described. Cells were labeled using ERα or ERβ antibodies along with prostate stem cell markers CD49f and TROP2 followed by triple channel FACS to quantify ERα+/ERβ+ cell numbers. To explore ERα, the benign human prostate stem cell line WPE with extremely low levels of ERα and ERβ, was stably transfected with a lentiviral-ERα expression vector. The human prostate cancer stem-like cell line HuSLC (ERβ++, ERα−) was utilized to interrogate ERb actions. Cells were exposed to 10 nM estradiol (E2) over a 15 to 60 minute time course +/− ICI 182,870 (ICI), an ERα/β antagonist. FACS analysis of day 7 PS cells labeled for ERα or ERβ revealed 66% of day 7 PS cells as ERα+ and 40% as ERβ+. Among ERα or ERβ positive PS cells, 4% were Trop2+/CD49fhigh (stem-like cells) and 10–12% were Trop2+/CD49fmedium (early stage progenitor cells). PS exposed to 10 nM E2 showed sequential phosphorylation of Src, Erk1/2, p38, Akt and NFκB (p65) over 60 minutes. Phosphorylation of up-and downstream targets (EGFR, Jnk, GSK 3α/β, p70 S6 kinase, PRAS40, MSK1/2) was also seen using a phospho-kinase array. Furthermore, phosphorylation of ERα at S167 was noted over 60 min of E2 exposure enabling enhancement of genomic ERα transactivational activity in a feed-forward manner. ICI attenuated Akt and Erk1/2 phosphorylation, confirming membrane bound ERs are involved in downstream signaling. E2 treatment of HuSLCs showed phosphorylation of Erk1/2 but not Akt, indicating that ERβ signals exclusively through the MAPK pathway in these cells. Conversely, E2 treatment of WPE-stem cells overexpressing ERα resulted in robust phosphorylation of Akt but lower levels of Erk1/2 phosphorylation suggesting that Akt activation may be more reliant on ERα signaling. To identify pathway specific roles, specific inhibitors were added to PS cultures. PS treated with LY294002 (Akt inhibitor) for 7 days attenuated the E2-mediated increase in PS number and size. Inhibition of the NFκB downstream of the Akt pathway by IKK VII (IKK inhibitor) blocked p65 phosphorylation, abrogated the E2-induced increase in stem cell symmetric self-renewal and blunted E2 stimulation of progenitor cell proliferation. Analysis of PS cyclin mRNA levels revealed a G1 arrest of progenitor cells upon IKK inhibition suggesting an essential role of NFκB in progenitor cell amplification. MAPK pathway inhibition with U0126(Erk1/2 inhibitor) resulted in an attenuation of the E2-mediated increase in PS number and size and an increase stem cell symmetric self-renewal suggesting that MAPK pathway activation promotes commitment to stem and progenitor cell expansion. Taken together, the present findings reveal that human prostate stem-progenitor cells express both ERα and ERβ which differentially activate different signaling cascades originating at the membrane. These signaling events may lead to unique downstream actions that influence prostate stem-progenitor cell proliferation as well as lineage commitment decisions.

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Are MSCs Stem Cells? Demonstration of in Vitro and in Vivo Stem Cell Properties of Highly Enriched linneg/CD45neg/CD271pos/CD140alow/Neg Mesenchymal Stromal Cells from Adult Human Bone Marrow

Blood ◽

10.1182/blood.v124.21.4374.4374 ◽

2014 ◽

Vol 124 (21) ◽

pp. 4374-4374

Author(s):

Roshanak Ghazanfari ◽

Hongzhe Li ◽

Dimitra Zacharaki ◽

Simón Méndez-Ferrer ◽

Stefan Scheding

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

Human Bone ◽

Human Bone Marrow ◽

Differentiation Potential ◽

Self Renewal ◽

Adult Human

Abstract Human bone marrow contains a rare population of non-hematopoietic mesenchymal stromal cells (BM-MSC) with multilineage differentiation capacity, which are essential constituents of the hematopoietic microenvironment. Self-renewal and differentiation are the two key properties of somatic stem cells, however, stem cell properties of human adult BM-MSC have not been demonstrated conclusively yet. We have previously shown that low/negative expression of PDGFRα on linneg/CD45neg/CD271pos cells identified a highly enriched population of primary BM-MSC in adult human bone marrow (Li et al. Blood, 2013, 122:3699). Based on this work, the current study aimed to investigate the in-vitro and in-vivo stem cell properties of this putative stromal stem cell population. The in-vitro clonogenic potential of freshly sorted human linneg/CD45neg/CD271pos/PDGFRlow/neg cells was evaluated by utilizing the CFU-F assay as well as the recently-developed mesensphere assay, which enables MSC amplification while preserving an immature phenotype (Isern et al, Cell Reports 2013, 30: 1714-24). Comparable colony frequencies were obtained with both assays (19.3 ± 2 and 17.5 ± 2.3 CFU-F and spheres per 100 plated cells, respectively, n=6, p=0.19). In order to test whether both assays identified the same population of clonogenic cells, colonies and spheres were replated under both conditions for up to three generations. The results showed comparable capacities of CFU-F and mesenspheres to form secondary and tertiary CFU-F and spheres. In-vitro self-renewal as indicated by increasing numbers of CFU-F and spheres (416.6 ± 431.7-fold and 49.5 ± 65.7-fold, respectively, n=3) was observed up to the third generation and decreased thereafter. The total number of generations was five (CFU-F) and six (spheres). In-vitro differentiation assays with both, CFU-F- and sphere-derived cells (tested until passage three) demonstrated tri-lineage differentiation potential (adipocytes, osteoblasts, chondrocytes). In addition, CFU-Fs and spheres had comparable surface marker profiles (CD73, CD90, CD105, and HLA-ABC positive; CD31, CD34 and HLA-DR negative), except for CD90, which was higher expressed on CFU-Fs. To investigate in-vivo self-renewal and differentiation potential of the putative stromal stem cells, linneg/CD45neg/CD271pos/PDGFRlow/neg -derived CFU-F and spheres were serially transplanted s.c into NSG mice. After 8 weeks, implants were harvested, human cells were FACS-isolated (CD90 and CD105 expression), and re-assayed under CFU-F and sphere conditions. Whereas in-vivo self-renewal of CFU-F could not be shown (111.5 ± 36 –fold decrease in total CFU-F numbers after primary transplantation, n=3), sphere self-renewal was clearly demonstrated by increased numbers of spheres after primary as well as secondary transplantation (1.13 ± 0.05 and 2.06 ± 0.26 –fold, respectively, n=3), which is remarkable given the fact that the number of recovered human cells is underestimated due to the isolation approach. Here, confirming GFP-marking experiments are ongoing. Finally, preliminary data indicate that linneg/CD45neg/CD271pos/PDGFRlow/neg –derived spheres display full in-vivo differentiation capacity in primary and secondary transplantations. Taken together, our data demonstrate - for the first time - that primary human linneg/CD45neg/CD271pos/PDGFRlow/neg cells meet stringent stem cell criteria, i.e. in-vitro and in-vivo self-renewal and differentiation. These findings answer the long-open question of the potential stem cell properties of adult human MSC and will enable to better understand the properties of native BM-MSC and their biological role in the bone marrow. Disclosures No relevant conflicts of interest to declare.

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Vascular Tissue Engineering: Polymers and Methodologies for Small Caliber Vascular Grafts

Frontiers in Cardiovascular Medicine ◽

10.3389/fcvm.2020.592361 ◽

2021 ◽

Vol 7 ◽

Author(s):

Bruna B. J. Leal ◽

Naohiro Wakabayashi ◽

Kyohei Oyama ◽

Hiroyuki Kamiya ◽

Daikelly I. Braghirolli ◽

...

Keyword(s):

Tissue Engineering ◽

Vascular Tissue ◽

Thrombus Formation ◽

Treatment Options ◽

Vascular Grafts ◽

Vascular Tissue Engineering ◽

Engineering Polymers ◽

Autologous Grafts

Cardiovascular disease is the most common cause of death in the world. In severe cases, replacement or revascularization using vascular grafts are the treatment options. While several synthetic vascular grafts are clinically used with common approval for medium to large-caliber vessels, autologous vascular grafts are the only options clinically approved for small-caliber revascularizations. Autologous grafts have, however, some limitations in quantity and quality, and cause an invasiveness to patients when harvested. Therefore, the development of small-caliber synthetic vascular grafts (<5 mm) has been urged. Since small-caliber synthetic grafts made from the same materials as middle and large-caliber grafts have poor patency rates due to thrombus formation and intimal hyperplasia within the graft, newly innovative methodologies with vascular tissue engineering such as electrospinning, decellularization, lyophilization, and 3D printing, and novel polymers have been developed. This review article represents topics on the methodologies used in the development of scaffold-based vascular grafts and the polymers used in vitro and in vivo.

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Neural crest–derived cells with stem cell features can be traced back to multiple lineages in the adult skin

The Journal of Cell Biology ◽

10.1083/jcb.200606062 ◽

2006 ◽

Vol 175 (6) ◽

pp. 1005-1015 ◽

Cited By ~ 224

Author(s):

Christine E. Wong ◽

Christian Paratore ◽

María T. Dours-Zimmermann ◽

Ariane Rochat ◽

Thomas Pietri ◽

...

Keyword(s):

Stem Cell ◽

Neural Crest ◽

Stem Cell Markers ◽

Self Renewal ◽

Neural Crest Stem Cell ◽

Stem And Progenitor Cells ◽

The Face ◽

Adult Skin ◽

Trunk Skin

Given their accessibility, multipotent skin-derived cells might be useful for future cell replacement therapies. We describe the isolation of multipotent stem cell–like cells from the adult trunk skin of mice and humans that express the neural crest stem cell markers p75 and Sox10 and display extensive self-renewal capacity in sphere cultures. To determine the origin of these cells, we genetically mapped the fate of neural crest cells in face and trunk skin of mouse. In whisker follicles of the face, many mesenchymal structures are neural crest derived and appear to contain cells with sphere-forming potential. In the trunk skin, however, sphere-forming neural crest–derived cells are restricted to the glial and melanocyte lineages. Thus, self-renewing cells in the adult skin can be obtained from several neural crest derivatives, and these are of distinct nature in face and trunk skin. These findings are relevant for the design of therapeutic strategies because the potential of stem and progenitor cells in vivo likely depends on their nature and origin.

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Atomic Force and Confocal Microscopic Studies of Collagen-Cell-Based Scaffolds for Vascular Tissue Engineering

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.15-17.83 ◽

2006 ◽

Vol 15-17 ◽

pp. 83-88 ◽

Cited By ~ 3

Author(s):

Luca Amadori ◽

Navneeta Rajan ◽

Simone Vesentini ◽

D. Mantovani

Keyword(s):

Tissue Engineering ◽

Smooth Muscle ◽

Smooth Muscle Cells ◽

Vascular Tissue ◽

Collagen Gel ◽

Muscle Cells ◽

Vascular Tissue Engineering ◽

Vascular Cells ◽

Atomic Force

Collagen is the most used naturally occurring scaffold material. It’s a structural protein ubiquitous among mammalian. The ability of collagen type I to host different cell phenotype in vitro and its low antigenecity in vivo are well known. However, the principal drawback of collagenbased materials consists in their low mechanical properties. For vascular tissue engineering this represents a major limit, as the aim is to mimic the structure of a native vessel, which is known to be resistant and viscoelastic. Moreover, vascular cells are known to be susceptible in vivo to reorganize the matrix in which they proliferate. Therefore, the aim of this project is to study the micro structural organization of collagen-based scaffolds, and to assess the interactions between collagen and smooth muscle cells during regeneration. This knowledge will then allow the development of appropriate strategies to tailor the microstructure of the scaffold and its properties. Smooth muscle cells (SMCs) were selected to study the interactions between cells and matrix during the proliferation. Atomic Force Microscopy (AFM) in dry state in tapping mode and Confocal Laser Scanning Microscopy (CLSM) in reflection mode were used to investigate the microstructure of the scaffold. For the former technique cells were seeded on top of the collagen gel after jellification, while for the latter, cells were embedded into the collagen gel and stained with Rhodamine. The contact points between matrix and cells were investigated, as well as the capacity of vascular cells to induce a structural reorganization of collagen fibrils in the scaffold.

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Engineering blood vessels and vascularized tissues: technology trends and potential clinical applications

Clinical Science ◽

10.1042/cs20180155 ◽

2019 ◽

Vol 133 (9) ◽

pp. 1115-1135 ◽

Cited By ~ 14

Author(s):

Prafulla Chandra ◽

Anthony Atala

Keyword(s):

Tissue Engineering ◽

Vascular Tissue ◽

Vascular Grafts ◽

Small Diameter ◽

Clinical Applications ◽

Angiogenic Factors ◽

Vascular Tissue Engineering ◽

Technology Trends

Abstract Vascular tissue engineering has the potential to make a significant impact on the treatment of a wide variety of medical conditions, including providing in vitro generated vascularized tissue and organ constructs for transplantation. Since the first report on the construction of a biological blood vessel, significant research and technological advances have led to the generation of clinically relevant large and small diameter tissue engineered vascular grafts (TEVGs). However, developing a biocompatible blood-contacting surface is still a major challenge. Researchers are using biomimicry to generate functional vascular grafts and vascular networks. A multi-disciplinary approach is being used that includes biomaterials, cells, pro-angiogenic factors and microfabrication technologies. Techniques to achieve spatiotemporal control of vascularization include use of topographical engineering and controlled-release of growth/pro-angiogenic factors. Use of decellularized natural scaffolds has gained popularity for engineering complex vascularized organs for potential clinical use. Pre-vascularization of constructs prior to implantation has also been shown to enhance its anastomosis after implantation. Host-implant anastomosis is a phenomenon that is still not fully understood. However, it will be a critical factor in determining the in vivo success of a TEVGs or bioengineered organ. Many clinical studies have been conducted using TEVGs, but vascularized tissue/organ constructs are still in the research & development stage. In addition to technical challenges, there are commercialization and regulatory challenges that need to be addressed. In this review we examine recent advances in the field of vascular tissue engineering, with a focus on technology trends, challenges and potential clinical applications.

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Intravital Imaging of Young and Aged Stem Cells in the Marrow of Long Bones: Visualizing Mammalian Stem Cell Behavior in Real-Time in Vivo

Blood ◽

10.1182/blood.v112.11.2418.2418 ◽

2008 ◽

Vol 112 (11) ◽

pp. 2418-2418

Author(s):

Anja Köhler ◽

Vince Schmithorst ◽

Marie-Dominique Filippi ◽

Marnie A. Ryan ◽

Deidre Daria ◽

...

Keyword(s):

Stem Cell ◽

Progenitor Cells ◽

Progenitor Cell ◽

Intravital Imaging ◽

Long Bones ◽

Self Renewal ◽

Hematopoietic Stem ◽

Close Vicinity ◽

Stem And Progenitor Cells

Abstract Hematopoiesis, the process in which blood cells are generated from hematopoietic stem and progenitor cells (HSPCs) is primarily confined to the bone cavities. The interactions of hematopoietic cells with stroma cells forming niches inside the bone cavities are central to hematopoiesis, as these regulate cell proliferation, self-renewal and differentiation. Hematopoietic cell/stroma interactions have thus been, in analogy to the immunological synapse, named stem/progenitor cell synapses. So far, visualization of the behavior of somatic stem and progenitor cells in an undisturbed in vivo environment has not been reported for the mammalian system and consequently, the cellular dynamics of stem, progenitor and differentiated cells in vivo are only poorly defined. We developed and performed intravital time-lapse 2-photon microscopy in the marrow of the long bones (tibia) of mice to study the behavior and dynamics of differentiated hematopoietic cells as well as HPCs and HSCs in close vicinity to the endosteum in vivo over time. We demonstrate that HPCs as well as HSCs reside in close vicinity to the endosteum, further supporting the notion of an endosteal stem cell niche, and that they are, in contrast to differentiated macrophages and dendritic cells, solitary and immobile. Both HPCs and HSCs occupy distinct positions relative to the endosteum and show cell protrusion movement consistent with an active stem/progenitor cell synapse. Lastly, we report that aged HSCs show increased protrusion movement and localize more distantly to the endosteum compared to young HSCs. In addition, aged HSCs present with reduced adhesion to stroma as well as reduced polarity upon adhesion in vitro, implying a connection between altered stem cell dynamics in vivo and stem cell aging. The intravital imaging technology developed might establish a basis for further delineating additional important questions in stem cell biology like cellular mechanisms of hematopoietic stem cell self-renewal and differentiation in the context of the stroma/niche in vivo.

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