High density micromass cultures of embryonic limb bud mesenchymal cells: An in vitro model of endochondral skeletal development

1999 ◽  
Vol 35 (5) ◽  
pp. 262-269 ◽  
Author(s):  
Maria Alice Mello ◽  
Rocky S. Tuan
Keyword(s):  
In Vitro Model ◽  
Skeletal Development ◽  
Mesenchymal Cells ◽  
High Density ◽  
Limb Bud ◽  
Embryonic Limb ◽  
Vitro Model

scholarly journals Establishment of an In Vitro Model to Study Mesenchymal Cells During Postnatal Lung Development

2020 ◽  
Author(s):  
D. Perez Bravo ◽  
J.A. Rodriguez Castillo ◽  
A.V. Cherian ◽  
A. Fysikopoulos ◽  
C. Samakovlis ◽  
...  
Keyword(s):  
Lung Development ◽  
In Vitro Model ◽  
Mesenchymal Cells ◽  
Vitro Model ◽  
Postnatal Lung Development

Expression and functional involvement of N-cadherin in embryonic limb chondrogenesis

Development ◽  
1994 ◽  
Vol 120 (1) ◽  
pp. 177-187 ◽  
Author(s):  
S.A. Oberlender ◽  
R.S. Tuan
Keyword(s):  
Pattern Formation ◽  
Cell Aggregation ◽  
Mesenchymal Cells ◽  
Limb Bud ◽  
Time Period ◽  
Functional Involvement ◽  
Embryonic Limb ◽  
Cellular Condensation

Cell adhesion molecules have been shown to be important mediators of morphogenesis and pattern formation. In this study, we have shown that N-cadherin is expressed in a specific spatiotemporal manner in the developing limb bud during chondrogenesis in vivo and in cultured limb mesenchyme in vitro. The time period of maximal expression of N-cadherin corresponds to the period of active cellular condensation, an event believed to be a necessary prerequisite for chondrogenic differentiation. To directly assess the functional involvement of N-cadherin in cellular condensation, we have examined the effects of perturbing N-cadherin activity on both cell aggregation and chondrogenesis using NCD-2, a rat monoclonal antibody directed against the binding region of N-cadherin. Non-immune rat IgG was used as a control. Our results show that functional N-cadherin is necessary for chondrogenesis to proceed both in vivo and in vitro. Limb mesenchymal cells exhibited characteristic Ca(2+)-dependent cell aggregation in suspension, which was inhibited in the presence of exogenous NCD-2. In micromass cultures of limb mesenchymal cells, NCD-2 inhibited overt chondrogenesis in a dose-dependent manner. Furthermore, NCD-2 inhibition of chondrogenesis in micromass cultures was time-dependent, suggesting that N-cadherin is crucially involved during the latter half of the first 24 hours of culture, a time period most likely corresponding to active cellular condensation. NCD-2 also significantly influenced limb development when injected into embryonic limb buds in vivo. In addition to significant inhibition of chondrogenesis and developmental delays, gross developmental deformities and perturbation of overall pattern formation were also observed. Taken together, these results demonstrate that N-cadherin is functionally required in mediating the cell-cell interactions among mesenchymal cells important for chondrogenesis in micromass culture in vitro and in the intact limb bud in vivo.

Embryonic Limb Mesenchyme Micromass Culture as an In Vitro Model for Chondrogenesis and Cartilage Maturation

2003 ◽  
pp. 359-375 ◽  
Author(s):  
Anthony M. DeLise ◽  
Emanuela Stringa ◽  
Wendy A. Woodward ◽  
Maria Alice Mello ◽  
Rocky S. Tuan
Keyword(s):  
In Vitro Model ◽  
Micromass Culture ◽  
Limb Mesenchyme ◽  
Embryonic Limb ◽  
Vitro Model ◽  
And Cartilage

scholarly journals A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells

2013 ◽  
Vol 304 (4) ◽  
pp. C289-C298 ◽  
Author(s):  
Xia Guo ◽  
Steven L. Stice ◽  
Nolan L. Boyd ◽  
Shi-You Chen
Keyword(s):  
Smooth Muscle ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
In Vitro Model ◽  
Embryonic Stem ◽  
Mesenchymal Cells ◽  
Dependent Manner ◽  
Vitro Model

The objective of this study was to develop a novel in vitro model for smooth muscle cell (SMC) differentiation from human embryonic stem cell-derived mesenchymal cells (hES-MCs). We found that hES-MCs were differentiated to SMCs by transforming growth factor-β (TGF-β) in a dose- and time-dependent manner as demonstrated by the expression of SMC-specific genes smooth muscle α-actin, calponin, and smooth muscle myosin heavy chain. Under normal growth conditions, however, the differentiation capacity of hES-MCs was very limited. hES-MC-derived SMCs had an elongated and spindle-shaped morphology and contracted in response to the induction of carbachol and KCl. KCl-induced calcium transient was also evident in these cells. Compared with the parental cells, TGF-β-treated hES-MCs sustained the endothelial tube formation for a longer time due to the sustained SMC phenotype. Mechanistically, TGF-β-induced differentiation was both Smad- and serum response factor/myocardin dependent. TGF-β regulated myocardin expression via multiple signaling pathways including Smad2/3, p38 MAPK, and PI3K. Importantly, we found that a low level of myocardin was present in mesoderm prior to SMC lineage determination, and a high level of myocardin was not induced until the differentiation process was initiated. Taken together, our study characterized a novel SMC differentiation model that can be used for studying human SMC differentiation from mesoderm during vascular development.

An in vitro model for investigation of vitamin A effects on wound healing

2021 ◽  
pp. 1-6
Author(s):  
Hoda Keshmiri Neghab ◽  
Mohammad Hasan Soheilifar ◽  
Gholamreza Esmaeeli Djavid
Keyword(s):  
Wound Healing ◽  
Endothelial Cell ◽  
Vitamin A ◽  
In Vitro Model ◽  
Cellular Proliferation ◽  
Endothelial Cell Migration ◽  
Normal Fibroblast ◽  
Anti Inflammatory ◽  
Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inflammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inflammation responses.

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