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Angiogenic Potential of Human Neonatal Foreskin Stromal Cells in the Chick Embryo Chorioallantoic Membrane Model.

Vishnubalaji R, Atteya M, Al-Nbaheen M, Oreffo RO, Aldahmash A, Alajez NM - Stem Cells Int (2015)

Bottom Line: When hNSSCs were seeded onto the top of the CAM, human von Willebrand factor (hVWF), CD31, smooth muscle actin (SMA), and factor XIIIa positive cells were observed in the chick endothelium.Interestingly, undifferentiated hNSSCs showed a propensity to differentiate towards ectoderm with indication of epidermal formation with cells positive for CD1a, CK5/6, CK19, FXIIIa, and S-100 cells, which warrant further investigation.Our findings imply a potential angiogenic role for hNSSCs ex vivo in the differentiated and undifferentiated state, with potential contribution to blood vessel formation and potential application in tissue regeneration and vascularization.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi Arabia.

ABSTRACT
Several studies have demonstrated the multipotentiality of human neonatal foreskin stromal cells (hNSSCs) as being able to differentiate into adipocytes and osteoblasts and potentially other cell types. Recently, we demonstrated that hNSSCs play a role during in vitro angiogenesis and appear to possess a capacity to differentiate into endothelial-like cells; however, their angiogenic potential within an ex vivo environment remains unclear. Current study shows hNSSCs to display significant migration potential in the undifferentiated state and high responsiveness in the in vitro wound healing scratch assay. When hNSSCs were seeded onto the top of the CAM, human von Willebrand factor (hVWF), CD31, smooth muscle actin (SMA), and factor XIIIa positive cells were observed in the chick endothelium. CAMs transplanted with endothelial-differentiated hNSSCs displayed a higher number of blood vessels containing hNSSCs compared to CAMs transplanted with undifferentiated hNSSCs. Interestingly, undifferentiated hNSSCs showed a propensity to differentiate towards ectoderm with indication of epidermal formation with cells positive for CD1a, CK5/6, CK19, FXIIIa, and S-100 cells, which warrant further investigation. Our findings imply a potential angiogenic role for hNSSCs ex vivo in the differentiated and undifferentiated state, with potential contribution to blood vessel formation and potential application in tissue regeneration and vascularization.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemical staining of CAM alone and CAM grafted with hNSSCs for endothelial-related markers. CAMs transplanted with undifferentiated and differentiated hNSSCs (10 days) were processed and stained using the indicated antibody. Brown colour indicates areas with positive staining for the respective cell marker (smooth muscle actin (SMA), factor XIIIa, CD31, and von Willebrand factor (vWF)). The human leukocyte antigen (HLA-ABC) was used to differentiate the engraftment of human cells from chick cells (Bar = 100 μm).
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fig4: Immunohistochemical staining of CAM alone and CAM grafted with hNSSCs for endothelial-related markers. CAMs transplanted with undifferentiated and differentiated hNSSCs (10 days) were processed and stained using the indicated antibody. Brown colour indicates areas with positive staining for the respective cell marker (smooth muscle actin (SMA), factor XIIIa, CD31, and von Willebrand factor (vWF)). The human leukocyte antigen (HLA-ABC) was used to differentiate the engraftment of human cells from chick cells (Bar = 100 μm).

Mentions: We have previously shown the ability of hNSSCs to differentiate into endothelial-like cells in vitro [8]; however, the ability of hNSSCs to participate in neovasculature ex vivo has not been addressed to date. To assess hNSSCs ex vivo differentiation potential, the pellets of hNSSCs undifferentiated or differentiated under endothelial-induction conditions were cultured on PTFE confetti for 2 days as shown in schematic diagram (Figures 3(a) and 3(b)). Subsequently, cell pellets with confetti were transplanted onto CAMs by making 2 cm2 window on ten-day-old chick embryos (Figure 3(c)); after a further ten days of growth, CAMs were imaged and assessed for vascular formation (Figures 3(d)–3(f)). CAMs transplanted with differentiated and undifferentiated hNSSCs exhibited significant blood vessel formation. The number of vessel branching points was significantly (P ≤ 0.05) higher in CAMs transplanted with differentiated hNSSCs (190 ± branching points) compared to CAMs transplanted with undifferentiated hNSSCs (110 ± branching points, Figure 3(g)). Control CAM cultures displayed the lowest number of branching points (70 ± branching points). Similarly, immunohistochemical staining revealed larger number of blood vessels in CAMs transplanted with differentiated hNSSCs compared to CAMs transplanted with undifferentiated hNSSCs (Figures 3(i) and 3(h), resp.). In order to assess the contribution of hNSSCs to CAM's neovasculature, immunohistochemical investigations of CAMs were undertaken and, as shown in Figure 4, the presence of a significant number of human cells within and in regions surrounding neovessels was observed. Cells positive for the following human angiogenic markers: vWF, CD31, SMA, and FXIIIa were observed, whereas the use of the human antibody against HLA-ABC indicated the engraftment of hNSSCs in chick CAM without notable cross reactivity with the chick cells (Figure 4). Concordant with data presented in Figure 3, CAMs transplanted with differentiated cells showed higher numbers of blood vessels compared to CAMs transplanted with undifferentiated hNSSCs.


Angiogenic Potential of Human Neonatal Foreskin Stromal Cells in the Chick Embryo Chorioallantoic Membrane Model.

Vishnubalaji R, Atteya M, Al-Nbaheen M, Oreffo RO, Aldahmash A, Alajez NM - Stem Cells Int (2015)

Immunohistochemical staining of CAM alone and CAM grafted with hNSSCs for endothelial-related markers. CAMs transplanted with undifferentiated and differentiated hNSSCs (10 days) were processed and stained using the indicated antibody. Brown colour indicates areas with positive staining for the respective cell marker (smooth muscle actin (SMA), factor XIIIa, CD31, and von Willebrand factor (vWF)). The human leukocyte antigen (HLA-ABC) was used to differentiate the engraftment of human cells from chick cells (Bar = 100 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4499640&req=5

fig4: Immunohistochemical staining of CAM alone and CAM grafted with hNSSCs for endothelial-related markers. CAMs transplanted with undifferentiated and differentiated hNSSCs (10 days) were processed and stained using the indicated antibody. Brown colour indicates areas with positive staining for the respective cell marker (smooth muscle actin (SMA), factor XIIIa, CD31, and von Willebrand factor (vWF)). The human leukocyte antigen (HLA-ABC) was used to differentiate the engraftment of human cells from chick cells (Bar = 100 μm).
Mentions: We have previously shown the ability of hNSSCs to differentiate into endothelial-like cells in vitro [8]; however, the ability of hNSSCs to participate in neovasculature ex vivo has not been addressed to date. To assess hNSSCs ex vivo differentiation potential, the pellets of hNSSCs undifferentiated or differentiated under endothelial-induction conditions were cultured on PTFE confetti for 2 days as shown in schematic diagram (Figures 3(a) and 3(b)). Subsequently, cell pellets with confetti were transplanted onto CAMs by making 2 cm2 window on ten-day-old chick embryos (Figure 3(c)); after a further ten days of growth, CAMs were imaged and assessed for vascular formation (Figures 3(d)–3(f)). CAMs transplanted with differentiated and undifferentiated hNSSCs exhibited significant blood vessel formation. The number of vessel branching points was significantly (P ≤ 0.05) higher in CAMs transplanted with differentiated hNSSCs (190 ± branching points) compared to CAMs transplanted with undifferentiated hNSSCs (110 ± branching points, Figure 3(g)). Control CAM cultures displayed the lowest number of branching points (70 ± branching points). Similarly, immunohistochemical staining revealed larger number of blood vessels in CAMs transplanted with differentiated hNSSCs compared to CAMs transplanted with undifferentiated hNSSCs (Figures 3(i) and 3(h), resp.). In order to assess the contribution of hNSSCs to CAM's neovasculature, immunohistochemical investigations of CAMs were undertaken and, as shown in Figure 4, the presence of a significant number of human cells within and in regions surrounding neovessels was observed. Cells positive for the following human angiogenic markers: vWF, CD31, SMA, and FXIIIa were observed, whereas the use of the human antibody against HLA-ABC indicated the engraftment of hNSSCs in chick CAM without notable cross reactivity with the chick cells (Figure 4). Concordant with data presented in Figure 3, CAMs transplanted with differentiated cells showed higher numbers of blood vessels compared to CAMs transplanted with undifferentiated hNSSCs.

Bottom Line: When hNSSCs were seeded onto the top of the CAM, human von Willebrand factor (hVWF), CD31, smooth muscle actin (SMA), and factor XIIIa positive cells were observed in the chick endothelium.Interestingly, undifferentiated hNSSCs showed a propensity to differentiate towards ectoderm with indication of epidermal formation with cells positive for CD1a, CK5/6, CK19, FXIIIa, and S-100 cells, which warrant further investigation.Our findings imply a potential angiogenic role for hNSSCs ex vivo in the differentiated and undifferentiated state, with potential contribution to blood vessel formation and potential application in tissue regeneration and vascularization.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh 11461, Saudi Arabia.

ABSTRACT
Several studies have demonstrated the multipotentiality of human neonatal foreskin stromal cells (hNSSCs) as being able to differentiate into adipocytes and osteoblasts and potentially other cell types. Recently, we demonstrated that hNSSCs play a role during in vitro angiogenesis and appear to possess a capacity to differentiate into endothelial-like cells; however, their angiogenic potential within an ex vivo environment remains unclear. Current study shows hNSSCs to display significant migration potential in the undifferentiated state and high responsiveness in the in vitro wound healing scratch assay. When hNSSCs were seeded onto the top of the CAM, human von Willebrand factor (hVWF), CD31, smooth muscle actin (SMA), and factor XIIIa positive cells were observed in the chick endothelium. CAMs transplanted with endothelial-differentiated hNSSCs displayed a higher number of blood vessels containing hNSSCs compared to CAMs transplanted with undifferentiated hNSSCs. Interestingly, undifferentiated hNSSCs showed a propensity to differentiate towards ectoderm with indication of epidermal formation with cells positive for CD1a, CK5/6, CK19, FXIIIa, and S-100 cells, which warrant further investigation. Our findings imply a potential angiogenic role for hNSSCs ex vivo in the differentiated and undifferentiated state, with potential contribution to blood vessel formation and potential application in tissue regeneration and vascularization.

No MeSH data available.


Related in: MedlinePlus