scholarly journals Type I collagen, fibrin and PuraMatrix matrices provide permissive environments for human endothelial and mesenchymal progenitor cells to form neovascular networks

2011 ◽  
Vol 5 (4) ◽  
pp. e74-e86 ◽  
Author(s):  
Patrick Allen ◽  
Juan Melero-Martin ◽  
Joyce Bischoff
Keyword(s):  
Progenitor Cells ◽  
Type I Collagen ◽  
Type I ◽  
Mesenchymal Progenitor Cells

Enrichment of Mesenchymal Progenitor Cells from Mouse Compact Bone.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4105-4105
Author(s):  
Brenton Short ◽  
Emer Clarke ◽  
Terry Thomas ◽  
Allen C. Eaves ◽  
Albertus W. Wognum ◽  
...  
Keyword(s):  
Stem Cells ◽  
Animal Model ◽  
Progenitor Cells ◽  
Compact Bone ◽  
Mesenchymal Cells ◽  
Type I ◽  
Mesenchymal Progenitor Cells ◽  
Hemopoietic Cells ◽  
Reproducible Method

Abstract Adult mammalian bone marrow (BM) contains at least two distinct stem cell populations; the stem cells of the hemopoietic lineage and a second population termed mesenchymal stem cells (MSC) whose function is to maintain the non-hemopoietic BM elements as well as skeletal homeostasis. MSC have been implicated as potential targets in a range of cellular therapies for treatment of defects of both the hemopoietic and skeletal systems, and as vehicles for gene therapy. In order to evaluate the potential of these cells in various therapies, a pre-clinical animal model in which both the biology and potential therapeutic applications of these cells can be assessed is of fundamental importance. The goal of the current study was to develop a robust and reproducible method for the isolation of MSC from murine hemopoietic tissues. Tibiae and femurs harvested from C57BL6/J mice were gently crushed with a pestle to release the marrow. The bone fragments were subsequently cut into small pieces with a scalpel and digested in a solution containing 3mg/ml Type I collagenase to yield a population of compact bone (CB) derived cells. Mesenchymal progenitor cells (MPC) were detected using an in vitro assay for fibroblast-colony forming cells (CFU-F). CB cells were plated at 1000 or 5000 cells per cm2 in complete MesenCult™ medium for 12 days, after which CFU-F-derived colonies were enumerated. We show that CFU-F are present at a significantly higher frequency in mouse CB than in the BM (433±225 vs 11.7±3.5 colonies/106 cells respectively, n=3). Based on these data we developed a simple and robust immunomagnetic selection method to highly enrich MPC from mouse CB by depleting essentially all nucleated hemopoietic cells (CD45+) and red blood cells (Ter119+) using magnetic particles and antibodies to CD45 and Ter119, respectively. Target CD45−Ter119− cells initially comprised 1.1±0.5% (n=9) of the total CB fraction as assayed by FACS. Following depletion, CD45−Ter119− cells comprised 74.5±16% (n=6) of the cells and were enriched 205 fold for CFU-F compared to the starting population, with a CFU-F frequency of 1 per 11 cells plated, and a total CFU-F recovery of 57.9 ± 18.5%. Analysis of CD45−Ter119−Sca-1+ cells, a phenotype previously shown to enrich for MPC, revealed that these cells were enriched 50 fold following depletion, from 0.53±0.5 to 26.5±8.23% (n=3). The enriched MPCs cultured at low O2 tension were devoid of hemopoietic contaminants at passage 1 and 2 as shown by lack of CD45, Ter119 and CD11b expression. The cultured CB-derived MPCs were capable of extensive in vitro proliferation and maintained the ability to differentiate into cells of the osteogenic, adipogenic and chondrogenic lineages. Furthermore, irradiated cultured mesenchymal cells supported long-term culture-initiating cells (LTC-IC) in 4-week cultures of Sca-1+ BM cells under limiting dilution conditions, at frequencies similar to those detected using irradiated primary BM feeders (i.e. 1 per 1600). These data provide a rapid, reproducible method by which multipotent mesenchymal cells devoid of contaminating hemopoietic cells can be readily obtained from limited numbers of mice to study the biology of MSC as well as the use of these cells as therapeutic agents in a preclinical animal model.

scholarly journals IL-1/TNF-αInflammatory and Anti-Inflammatory Synchronization Affects Gingival Stem/Progenitor Cells’ Regenerative Attributes

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Fan Zhang ◽  
Misi Si ◽  
Huiming Wang ◽  
Mohamed K. Mekhemar ◽  
Christof E. Dörfer ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Osteogenic Differentiation ◽  
Progenitor Cells ◽  
Type I Collagen ◽  
Basic Medium ◽  
Control Group ◽  
Type I ◽  
Alizarin Red ◽  
Cell Numbers ◽  
Anti Inflammatory

Cytokines play major roles in tissue destruction/repair. The present study investigates proliferative and osteogenic differentiation potentials of gingival mesenchymal stem/progenitor cells (G-MSCs), influenced by IL-1/TNF-αinflammatory/anti-inflammatory conditions. Human G-MSCs were isolated, characterized, and cultured in basic medium (control group, M1), in basic medium with IL-1β, TNF-α, and IFN-γ(inflammatory group, M2) and with IL-1ra/TNF-αi added to M2 (anti-inflammatory group, M3). MTT tests at days 1, 3, and 7 and CFU assay at day 12 were conducted. Osteogenic differentiation was analyzed by bone-specific transcription factors (RUNX2), alkaline phosphatase (ALP), type I collagen (Col-I), osteopontin (OPN), and osteonectin (ON) expression at days 1, 3, 7, and 14 and Alizarin red staining at day 14. At day 3, the control group showed the highest cell numbers. At day 7, cell numbers in inflammatory and anti-inflammatory group outnumbered the control group. At day 12, CFUs decreased in the inflammatory and anti-inflammatory groups, with altered cellular morphology. The anti-inflammatory group demonstrated elevated bone-specific transcription factors at 14 days. After 14 days of osteogenic induction, calcified nodules in the anti-inflammatory group were higher compared to control and inflammatory groups. For regeneration, initial inflammatory stimuli appear essential for G-MSCs’ proliferation. With inflammatory persistence, this positive effect perishes and is followed by a short-term stimulatory one on osteogenesis. At this stage, selective anti-inflammatory intervention could boost G-MSCs’ differentiation.

scholarly journals Syndecan-4 Functionalization of Tissue Regeneration Scaffolds Improves Interaction with Endothelial Progenitor Cells

2021 ◽  
Author(s):  
Harleigh Warner ◽  
Yidi Wu ◽  
William D Wagner
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Tissue Regeneration ◽  
Binding Sites ◽  
Type I Collagen ◽  
Type I ◽  
Endothelial Progenitor ◽  
Hybrid Scaffold ◽  
Scaffold Materials

Abstract Key to most implanted cell free scaffolds for tissue regeneration is the ability to sequester and retain undifferentiated mesenchymal stem cells at the repair site. In this report, syndecan-4, a heparan sulfate containing proteoglycan, was investigated as a unique molecule for use in scaffold functionalization. An electrospun hybrid scaffold comprised of poly (glycerol) sebacate (PGS), silk fibroin and type I collagen (PFC) was used as a model scaffold to develop a procedure and test the hypothesis that functionalization would result in increased scaffold binding of endothelial progenitor cells (EPCs). For these studies both Syndecan-4 and stromal derived factor-1α (SDF-1α) were used in functionalization PFC. Syndecan-4 functionalized PFC bound 4.8 fold more SDF-1α compared to nonfunctionalized PFC. Binding was specific as determined by heparin displacement studies. After culture for 7 days, significantly, more EPCs were detected on PFC scaffolds having both syndecan-4 and SDF-1α compared to scaffolds of PFC with only syndecan-4, or PFC adsorbed with SDF-1α, or PFC alone. Taken together, this study demonstrates that EPCs can be bound to and significantly expanded on PFC material through syndecan-4 mediated growth factor binding. Syndecan-4 with a multiplicity of binding sites has the potential to functionalize and expand stem cells on a variety of scaffold materials for use in tissue regeneration.

scholarly journals Differential Production of Cartilage ECM in 3D Agarose Constructs by Equine Articular Cartilage Progenitor Cells and Mesenchymal Stromal Cells

2020 ◽  
Vol 21 (19) ◽  
pp. 7071
Author(s):  
Stefanie Schmidt ◽  
Florencia Abinzano ◽  
Anneloes Mensinga ◽  
Jörg Teßmar ◽  
Jürgen Groll ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Progenitor Cells ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Cell Types ◽  
Type I ◽  
Cartilage Engineering ◽  
Hypoxic Conditions

Identification of articular cartilage progenitor cells (ACPCs) has opened up new opportunities for cartilage repair. These cells may be used as alternatives for or in combination with mesenchymal stromal cells (MSCs) in cartilage engineering. However, their potential needs to be further investigated, since only a few studies have compared ACPCs and MSCs when cultured in hydrogels. Therefore, in this study, we compared chondrogenic differentiation of equine ACPCs and MSCs in agarose constructs as monocultures and as zonally layered co-cultures under both normoxic and hypoxic conditions. ACPCs and MSCs exhibited distinctly differential production of the cartilaginous extracellular matrix (ECM). For ACPC constructs, markedly higher glycosaminoglycan (GAG) contents were determined by histological and quantitative biochemical evaluation, both in normoxia and hypoxia. Differential GAG production was also reflected in layered co-culture constructs. For both cell types, similar staining for type II collagen was detected. However, distinctly weaker staining for undesired type I collagen was observed in the ACPC constructs. For ACPCs, only very low alkaline phosphatase (ALP) activity, a marker of terminal differentiation, was determined, in stark contrast to what was found for MSCs. This study underscores the potential of ACPCs as a promising cell source for cartilage engineering.

scholarly journals 4485 Silicone Implant Shells Increase the Rate of Proliferation of Patient-Derived BIA-ALCL Cells but Not Primary T Cells in an Engineered Biomimetic Breast Platform

2020 ◽  
Vol 4 (s1) ◽  
pp. 113-113
Author(s):  
Ishani Premaratne ◽  
Matthew Wright ◽  
Mariam Gadjiko ◽  
Daniel Lara ◽  
Arash Samadi ◽  
...  
Keyword(s):  
T Cells ◽  
Cell Count ◽  
Progenitor Cells ◽  
Type I Collagen ◽  
Breast Implant ◽  
Stromal Vascular Fraction ◽  
Large Cell ◽  
Silicone Implant ◽  
Seeding Density ◽  
Type I

OBJECTIVES/GOALS: We use a tissue engineered, biomimetic, 3D model to study the pathogenesis of breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) by comparing the effect of silicone implant shell on proliferation of patient-derived BIA-ALCL to its precursor T cells within the breast microenvironment. METHODS/STUDY POPULATION: Patient-derived breast tissue was processed for component adipocytes, ductal organoids, and stromal vascular fraction. These were suspended within 50 µl of 0.3% type I collagen matrix to which was added 200,000 cells/mL of either patient-derived BIA-ALCL cells or T progenitor cells. These were then plated into 6mm wells. As a control, both BIA-ALCL cells and T progenitor cells were suspended within type I collagen alone at the same seeding density without breast components. Before plating, wells were lined circumferentially with either textured, smooth, or no implant shell. These were 1cm by 2cm pieces dissected from the whole implant. Wells were imaged using confocal microscopy over 8 days. RESULTS/ANTICIPATED RESULTS: Unstimulated T progenitor cell count showed no significant increase in any of the conditions tested. The change in cell count over 8 days was 3.85% in each condition (p = 0.3352). A Tukey’s multiple comparison test comparing each condition revealed no significant increase in cell count over 8 days for all six conditions. Notably, our previous studies have shown proliferation of BIA-ALCL cells to be significantly more robust in the biomimetic platform compared to collagen-only groups, regardless of implant shell type (p < 0.01). BIA-ALCL cells grew nearly 30% faster in textured and smooth shell biomimetic groups compared to biomimetic wells lacking implant shell. DISCUSSION/SIGNIFICANCE OF IMPACT: Towards elucidating BIA-ALCL’s etiopathology, we show that silicone implant shell has a significant effect on proliferation of BIA-ALCL cells, but not their precursor T cells. If breast implant silicone shell is not a sufficient stimulus for T cell proliferation, co-stimulatory factors are required.

scholarly journals The tyrosine kinase receptors Ron and Sea control "scattering" and morphogenesis of liver progenitor cells in vitro.

1996 ◽  
Vol 7 (4) ◽  
pp. 495-504 ◽  
Author(s):  
E Medico ◽  
A M Mongiovi ◽  
J Huff ◽  
M A Jelinek ◽  
A Follenzi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Tyrosine Kinase ◽  
Progenitor Cells ◽  
Type I Collagen ◽  
Growth Factor Receptor ◽  
Biological Response ◽  
Tyrosine Kinase Receptors ◽  
Type I ◽  
Macrophage Stimulating Protein

The mammalian RON and the avian sea genes encode tyrosine kinase receptors of poorly characterized biological functions. We recently identified macrophage-stimulating protein as the ligand for Ron; no ligand has yet been found for Sea. In this work we investigated the biological response to macrophage-stimulating protein in mouse liver progenitor cells expressing Ron. These cells were also transfected with a chimeric cDNA encoding the cytoplasmic domain of Sea, fused to the extracellular domain of Trk (nerve growth factor receptor). In the presence of nanomolar concentrations of the respective ligands, both receptors induced cell "scattering", extracellular matrix invasion, and DNA synthesis. When liver progenitor cells were grown in a tri-dimensional type-I collagen matrix, ligand-induced stimulation of either Ron or Sea induced sprouting of branched cell cords, evolving into ductular-like tubules. The motogenic, mitogenic, and morphogenic responses were also elicited by triggering the structurally related hepatocyte growth factor receptor (Met) but not epidermal growth factor or platelet-derived growth factor receptors. These data show that Ron, Sea, and Met belong to a receptor subfamily that elicits a distinctive biological response in epithelial cells.

Mesenchymal Progenitor Cells Communicate via Alpha and Beta Integrins with a Three-Dimensional Collagen Type I Matrix

2006 ◽  
Vol 182 (3-4) ◽  
pp. 143-154 ◽  
Author(s):  
Leslie Heckmann ◽  
Jörg Fiedler ◽  
Thomas Mattes ◽  
Rolf E. Brenner
Keyword(s):  
Progenitor Cells ◽  
Collagen Type ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Type I ◽  
Mesenchymal Progenitor Cells

scholarly journals Three-Dimensional Modeling of the Structural Microenvironment in Post-Traumatic War Wounds

2021 ◽  
Author(s):  
Gregory T. Christopherson ◽  
Jaira F. de Vasconcellos ◽  
John C. Dunn ◽  
Daniel W. Griffin ◽  
Patrick E. Jones ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Structural Changes ◽  
Type I Collagen ◽  
Three Dimensional ◽  
Specific Treatment ◽  
High Energy ◽  
Type I ◽  
Differentiation Potential ◽  
War Wounds ◽  
Post Traumatic

Abstract BACKGROUND: The development of post-traumatic heterotopic ossification (HO) is a common, undesirable sequela in patients with high-energy (war-related) extremity injuries. While inflammatory and osteoinductive signaling pathways are known to be involved in the development and progression of post-traumatic HO, features of the structural microenvironment within which the ectopic bone begins to form remain poorly understood. Thus, increasing our knowledge of molecular and structural changes within the healing wound may help elucidate the pathogenesis of post-traumatic HO and aid in the development of specific treatment and/or prevention strategies. METHODS: In this study, we performed high-resolution microscopy and biochemical analysis of tissues obtained from traumatic war wounds to characterize changes in the structural microenvironment. In addition, using an electrospinning approach, we modeled this microenvironment to reconstitute a three-dimensional type I collagen scaffold with non-woven, randomly oriented nanofibers where we evaluated the performance of primary mesenchymal progenitor cells. RESULTS: We found that traumatic war wounds are characterized by a disorganized, densely fibrotic collagen I matrix that influences progenitor cells adhesion, proliferation and osteogenic differentiation potential. CONCLUSION: Altogether, these results suggest that the structural microenvironment present in traumatic war wounds has the potential to contribute to the development of post-traumatic HO. Our findings may support novel treatment strategies directed towards modifying the structural microenvironment after traumatic injury.

scholarly journals Type I and II collagen regulation of chondrogenic differentiation by mesenchymal progenitor cells

2005 ◽  
Vol 23 (2) ◽  
pp. 446-453 ◽  
Author(s):  
C. W. Chen ◽  
Y. H. Tsai ◽  
W. P. Deng ◽  
S. N. Shih ◽  
C. L. Fang ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Chondrogenic Differentiation ◽  
Type I ◽  
Mesenchymal Progenitor Cells

Efficient induction of neural progenitor cells from human ESC/iPSCs on Type I Collagen

2021 ◽  
Author(s):  
Pengfei Liu ◽  
Shubin Chen ◽  
Yaofeng Wang ◽  
Xiaoming Chen ◽  
Yiping Guo ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Neural Progenitor Cells ◽  
Type I Collagen ◽  
Neural Progenitor ◽  
Type I ◽  
Efficient Induction
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