Labeling and Qualification of Endothelial Progenitor Cells for Tracking in Tissue Engineering: An <i>in Vitro</i> Study

Thebaud, Noëlie; Aussel, Audrey; Siadous, Robin; Toutain, Jérôme; Bareille, Reine; Montembault, Alexandra; David, Laurent; Bordenave, Laurence

doi:10.5301/ijao.5000405

Cited by 15 publications

(12 citation statements)

References 24 publications

(48 reference statements)

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“…Samples were treated anonymously. HUVECs were transduced with a vector containing the RFP gene under the control of the EF1a promoter following a procedure similar to the one previously described [24]. RFP-labeled HUVECs were cultured in Iscove's Modified Dulbecco's Medium (IMDM, Gibco, Paisley, Scotland, UK) containing 20% fetal bovine serum (FBS, GE Healthcare, Pasching, Austria), 20 μg ml −1 endothelial cell growth supplement, 90 μg ml −1 heparin (ECGS/H 0.4% (v/v); PromoCell, Heidelberg, Germany) and 10% endothelial cell growth medium-2 (EGM-2 MV, Lonza, Walkersville, MD, USA), in a controlled atmosphere (5% CO 2 , 95% relative humidity (RH), 37 °C-incubator (Binder, Germany)).…”

Section: Methodsmentioning

confidence: 99%

In situ prevascularization designed by laser-assisted bioprinting: effect on bone regeneration

et al. 2019

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Vascularization plays a crucial role in bone formation and regeneration process. Development of a functional vasculature to improve survival and integration of tissue-engineered bone substitutes remains a major challenge. Biofabrication technologies, such as bioprinting, have been introduced as promising alternatives to overcome issues related to lack of prevascularization and poor organization of vascular networks within the bone substitutes. In this context, this study aimed at organizing endothelial cells in situ, in a mouse calvaria bone defect, to generate a prevascularization with a defined architecture, and promote in vivo bone regeneration. Laser-assisted bioprinting (LAB) was used to pattern Red Fluorescent Protein-labeled endothelial cells into a mouse calvaria bone defect of critical size, filled with collagen containing mesenchymal stem cells and vascular endothelial growth factor. LAB technology allowed safe and controlled in vivo printing of different cell patterns. In situ printing of endothelial cells gave rise to organized microvascular networks into bone defects. At two months, vascularization rate (vr) and bone regeneration rate (br) showed statistically significant differences between the ‘random seeding’ condition and both ‘disc’ pattern (vr = +203.6%; br = +294.1%) and ‘crossed circle’ pattern (vr = +355%; br = +602.1%). These results indicate that in vivo LAB is a valuable tool to introduce in situ prevascularization with a defined configuration and promote bone regeneration.

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Section: Methodsmentioning

confidence: 99%

In situ prevascularization designed by laser-assisted bioprinting: effect on bone regeneration

et al. 2019

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“…Cells were expanded in Iscove's Modified Dulbecco's Medium (IMDM; Gibco, Thermo Fisher Scientific) containing 20% FBS, 12 μ g/mL endothelial cell growth supplement, and 90 μ g/mL heparin (ECGS/H 0.4% (v/v); PromoCell, Heidelberg, Germany) in gelatin-coated (0.2%; Sigma-Aldrich, Saint Quentin Fallavier, France) cell-culture flasks. Passage-1 cells were transduced with a lentiviral vector codding for the tdTomato fluorescent protein [18, 19]. Transduced and untransduced cells were used as stated for each experiment.…”

Section: Methodsmentioning

confidence: 99%

Patterning of Endothelial Cells and Mesenchymal Stem Cells by Laser-Assisted Bioprinting to Study Cell Migration

Bourget

Kérourédan

Medina

et al. 2016

BioMed Research International

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Tissue engineering of large organs is currently limited by the lack of potent vascularization in vitro. Tissue-engineered bone grafts can be prevascularized in vitro using endothelial cells (ECs). The microvascular network architecture could be controlled by printing ECs following a specific pattern. Using laser-assisted bioprinting, we investigated the effect of distance between printed cell islets and the influence of coprinted mesenchymal cells on migration. When printed alone, ECs spread out evenly on the collagen hydrogel, regardless of the distance between cell islets. However, when printed in coculture with mesenchymal cells by laser-assisted bioprinting, they remained in the printed area. Therefore, the presence of mesenchymal cell is mandatory in order to create a pattern that will be conserved over time. This work describes an interesting approach to study cell migration that could be reproduced to study the effect of trophic factors.

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“…Most cell tracking work has centered on imaging modalities such as MRI, radionuclide imaging, and optical imaging. However, X-ray computed tomography (CT) is an emerging method for cell tracking that has several strengths such as high spatial and temporal resolution, and excellent quantitative capabilities [2-4,11,12].…”

Section: Discussionmentioning

confidence: 99%

Cell Versus Chemokine Therapy Effects on Cell Mobilization to Chronically Dysfunctional Urinary Sphincters of Nonhuman Primates

et al. 2018

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PurposeA major question remaining in approaches to tissue engineering and organ replacement is the role of native mobilized native cells in the regeneration process of damaged tissues and organs. The goal of this study was to compare the cell mobilizing effects of the chemokine CXCL12 and cell therapy on the urinary sphincter of nonhuman primates (NHP) with chronic intrinsic urinary sphincter dysfunction.MethodsEither autologous lenti-M-cherry labeled skeletal muscle precursor cells (skMPCs) or CXCL12 were injected directly into the sphincter complex of female NHPs with or without surgery-induced chronic urinary sphincter dysfunction (n=4/treatment condition). All monkeys had partial bone marrow transplantation with autologous lenti-green fluorescent protein (GFP) bone marrow cells prior to treatment. Labeled cells were identified, characterized and quantified using computer-assisted immunohistochemistry 6 months posttreatment.ResultsGFP-labeled bone marrow cells (BMCs) were identified in the bone marrow and both BMCs and skMPCs were found in the urinary sphincter at 6-month postinjection. BMCs and skMPCs were present in the striated muscle, smooth muscle, and lamina propria/urothelium of the sphincter tissue. Sphincter injury increased the sphincter content of BMCs when analyzed 6-month postinjection. CXCL12 treatment, but not skMPCs, increased the number of BMCs in all layers of the sphincter complex (P<0.05). CXCL12 only modestly (P=0.15) increased the number of skMPCs in the sphincter complex.ConclusionsThis dual labeling methodology now provides us with the tools to measure the relative number of locally injected cells versus bone marrow transplanted cells. The results of this study suggest that CXCL12 promotes mobilization of cells to the sphincter, which may contribute more to sphincter regeneration than injected cells.

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Labeling and Qualification of Endothelial Progenitor Cells for Tracking in Tissue Engineering: An in Vitro Study

Cited by 15 publications

References 24 publications

In situ prevascularization designed by laser-assisted bioprinting: effect on bone regeneration

In situ prevascularization designed by laser-assisted bioprinting: effect on bone regeneration

Patterning of Endothelial Cells and Mesenchymal Stem Cells by Laser-Assisted Bioprinting to Study Cell Migration

Cell Versus Chemokine Therapy Effects on Cell Mobilization to Chronically Dysfunctional Urinary Sphincters of Nonhuman Primates

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