Investigating the role of hematopoietic stem and progenitor cells in regulating the osteogenic differentiation of mesenchymal stem cells in vitro

Liao, Jiehong; Hammerick, Kyle E.; Challen, Grant A.; Goodell, Margaret A.; Kasper, F. Kurtis; Mikos, Antonios G.

doi:10.1002/jor.21436

Cited by 26 publications

(29 citation statements)

References 36 publications

(54 reference statements)

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“…However, unlike previous studies, there was also an increase in late osteogenic markers (calcium) compared with the control group (osteogenic). Importantly, and in contrast to previous studies, 37,[40][41][42][43][44][46][47][48][49]69 this increase in ALP and calcium content was achieved in a 3D scaffoldless construct, without the use of any osteogenic supplements. When compared with the calcium content results, 49 the increase in calcium content in the coculture group with added MSCs/ HUVECs, but no osteogenic supplements, was significantly higher than in the same group cultured in osteogenic supplements as reported by Freeman et al 49 There was significantly higher calcium content in the group cultured with osteogenic supplements than in the group without Quantitative analysis of cross-sectional area of the rudimentary vessels present in the aggregates.…”

Section: Discussioncontrasting

confidence: 89%

“…Moreover, similar to previous studies, 37,[40][41][42][43][44][46][47][48][49]69 the coculture group with both HUVECs and MSCs added had significantly higher ALP production than the control groups (osteogenic and noncoculture group) after 3 weeks of coculture. However, unlike previous studies, there was also an increase in late osteogenic markers (calcium) compared with the control group (osteogenic).…”

Section: Discussionsupporting

confidence: 88%

“…Previous in vitro studies have shown that direct coculture of MSCs or osteoblasts with endothelial cells can upregulate production of the early osteogenic marker ALP, [36][37][38][39] without the presence of osteogenic supplements. Other studies have investigated whether coculture of MSCs and endothelial cells can increase ALP production in three-dimensional (3D) polymer scaffolds 40,41 or 3D cellular aggregates, [42][43][44][45] but the majority of these studies were conducted in the presence of osteogenic growth supplements. [42][43][44][45] The osteogenic potential of MSC/chondrocyte or osteoblast/ chondrocyte cocultures has been variable and inconclusive in both two-dimensional (2D) and 3D cultures.…”

mentioning

confidence: 99%

“…Other studies have investigated whether coculture of MSCs and endothelial cells can increase ALP production in three-dimensional (3D) polymer scaffolds 40,41 or 3D cellular aggregates, [42][43][44][45] but the majority of these studies were conducted in the presence of osteogenic growth supplements. [42][43][44][45] The osteogenic potential of MSC/chondrocyte or osteoblast/ chondrocyte cocultures has been variable and inconclusive in both two-dimensional (2D) and 3D cultures. [46][47][48] One study investigated the effect of coculture of hMSCs and chondrocytes, without the use of osteogenic supplements, and found that there was no ALP production/ expression in 3D aggregate culture.…”

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confidence: 99%

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Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Freeman

Stevens

Owens

et al. 2016

Tissue Engineering Part A

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In vitro bone regeneration strategies that prime mesenchymal stem cells (MSCs) with chondrogenic factors, to mimic aspects of the endochondral ossification process, have been shown to promote mineralization and vascularization by MSCs both in vitro and when implanted in vivo. However, these approaches required the use of osteogenic supplements, namely dexamethasone, ascorbic acid, and b-glycerophosphate, none of which are endogenous mediators of bone formation in vivo. Rather MSCs, endothelial progenitor cells, and chondrocytes all reside in proximity within the cartilage template and might paracrineally regulate osteogenic differentiation. Thus, this study tests the hypothesis that an in vitro bone regeneration approach that mimics the cellular niche existing during endochondral ossification, through coculture of MSCs, endothelial cells, and chondrocytes, will obviate the need for extraneous osteogenic supplements and provide an alternative strategy to elicit osteogenic differentiation of MSCs and mineral production. The specific objectives of this study were to (1) mimic the cellular niche existing during endochondral ossification and (2) investigate whether osteogenic differentiation could be induced without the use of any external growth factors. To test the hypothesis, we evaluated the mineralization and vessel formation potential of (a) a novel methodology involving both chondrogenic priming and the coculture of human umbilical vein endothelial cells (HUVECs) and MSCs compared with (b) chondrogenic priming of MSCs alone, (c) addition of HUVECs to chondrogenically primed MSC aggregates, (d-f) the same experimental groups cultured in the presence of osteogenic supplements and (g) a noncoculture group cultured in the presence of osteogenic growth factors alone. Biochemical (DNA, alkaline phosphatase [ALP], calcium, CD31 + , vascular endothelial growth factor [VEGF]), histological (alcian blue, alizarin red), and immunohistological (CD31 + ) analyses were conducted to investigate osteogenic differentiation and vascularization at various time points (1, 2, and 3 weeks). The coculture methodology enhanced both osteogenesis and vasculogenesis compared with osteogenic differentiation alone, whereas osteogenic supplements inhibited the osteogenesis and vascularization (ALP, calcium, and VEGF) induced through coculture alone. Taken together, these results suggest that chondrogenic and vascular priming can obviate the need for osteogenic supplements to induce osteogenesis of human MSCs in vitro, while allowing for the formation of rudimentary vessels.

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Section: Discussioncontrasting

confidence: 89%

Section: Discussionsupporting

confidence: 88%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Freeman

Stevens

Owens

et al. 2016

Tissue Engineering Part A

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“…In particular, depending on its composition, ECM retains the capability of either maintaining cell stemness 11 or conversely enhancing the osteogenic potential of MSCs. 12,13 Moreover, the initial cellularity of the samples is another important, but often underestimated factor, further affecting the development of the MSC/scaffold constructs. 14,15 However, neither has the role performed by transient populations been deepened so far, nor have celldynamic biomimetic osteogenic niches been investigated in TE studies.…”

mentioning

confidence: 99%

Growing Bone Tissue-Engineered Niches with Graded Osteogenicity: AnIn VitroMethod for Biomimetic Construct Assembly

Serino

D’Alessandro

Moscato

et al. 2013

Tissue Engineering Part C: Methods

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The traditional bone tissue-engineering approach exploits mesenchymal stem cells (MSCs) to be seeded once only on three-dimensional (3D) scaffolds, hence, differentiated for a certain period of time and resulting in a homogeneous osteoblast population at the endpoint. However, after achieving terminal osteodifferentiation, cell viability is usually markedly compromised. On the other hand, naturally occurring osteogenesis results from the coexistence of MSC progenies at distinct differentiative stages in the same microenvironment. This diversification also enables long-term viability of the mature tissue. We report an easy and tunable in vitro method to engineer simple osteogenic cell niches in a biomimetic fashion. The niches were grown via periodic reseeding of undifferentiated MSCs on MSC/scaffold constructs, the latter undergoing osteogenic commitment. Timefractioning of the seeded cell number during differentiation time of the constructs allowed graded osteogenic cell populations to be grown together on the same scaffolds (i.e., not only terminally differentiated osteoblasts). In such cell-dynamic systems, the overall differentiative stage of the constructs could also be tuned by varying the cell density seeded at each inoculation. In this way, we generated two different biomimetic niche models able to host good reservoirs of preosteoblasts and other osteoprogenitors after 21 culture days. At that time, the niche type resulting in 40.8% of immature osteogenic progenies and only 59.2% of mature osteoblasts showed a calcium content comparable to the constructs obtained with the traditional culture method (i.e., 100.03 -29.30 vs. 78.51 -28.50 pg/cell, respectively; p = not significant), the latter colonized only by fully differentiated osteoblasts showing exhausted viability. This assembly method for tissue-engineered constructs enabled a set of important parameters, such as viability, colonization, and osteogenic yield of the MSCs to be balanced on 3D scaffolds, thus achieving biomimetic in vitro models with graded osteogenicity, which are more complex and reliable than those currently used by tissue engineers.

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Microfluidic Generation of Gradient Hydrogels to Modulate Hematopoietic Stem Cell Culture Environment

Mahadik

Wheeler

Skertich

et al. 2013

Adv Healthcare Materials

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The bone marrow provides spatially and temporally variable signals that impact the behavior of hematopoietic stem cells (HSCs). While multiple biomolecular signals and bone marrow cell populations have been proposed as key regulators of HSC fate, new tools are required to probe their importance and mechanisms of action. Here, a novel method based on a microfluidic mixing platform to create small volume, 3D hydrogel constructs containing overlapping patterns of cell and matrix constituents inspired by the HSC niche is described. This approach is used to generate hydrogels containing opposing gradients of fluorescent microspheres, MC3T3-E1 osteoblasts, primary murine hematopoietic stem and progenitor cells (HSPCs), and combinations thereof in a manner independent of hydrogel density and cell/particle size. Three different analytical methods are described to characterize local properties of these hydrogels at multiple scales: 1) whole construct fluorescent analysis; 2) multi-photon imaging of individual cells within the construct; 3) retrieval of discrete sub-regions from the hydrogel post-culture. The approach reported here allows the creation of stable gradients of cell and material cues within a single, optically translucent 3D biomaterial to enable a range of investigations regarding how microenvironmental signals impact cell fate.

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Investigating the role of hematopoietic stem and progenitor cells in regulating the osteogenic differentiation of mesenchymal stem cells in vitro

Cited by 26 publications

References 36 publications

Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Osteogenic Differentiation of Mesenchymal Stem Cells by Mimicking the Cellular Niche of the Endochondral Template

Growing Bone Tissue-Engineered Niches with Graded Osteogenicity: AnIn VitroMethod for Biomimetic Construct Assembly

Microfluidic Generation of Gradient Hydrogels to Modulate Hematopoietic Stem Cell Culture Environment

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