Autologous human plasma in stem cell culture and cryopreservation in the creation of a tissue-engineered vascular graft Ping Zhang, PhD, Aleksandra Policha,

Slides:

Advertisements

Similar presentations

Bisphenol A at environmentally relevant doses induces cyclooxygenase-2 expression and promotes invasion of human mesenchymal stem cells derived from uterine.

Advertisements

Human adult endothelial cell growth in culture

Vascular endothelial growth factor-C derived from CD11b+ cells induces therapeutic improvements in a murine model of hind limb ischemia Go Kuwahara,

Local lentiviral short hairpin RNA silencing of CCR2 inhibits vein graft thickening in hypercholesterolemic apolipoprotein E3-Leiden mice Daniël Eefting,

Angiogenic properties of sustained release platelet-rich plasma: Characterization in-vitro and in the ischemic hind limb of the mouse Shyamal Chandra.

Yong-An Lee, Ph. D. , Yong-Hee Kim, B. S. , Seung-Jung Ha, B. S

High mobility group box 1 promotes endothelial cell angiogenic behavior in vitro and improves muscle perfusion in vivo in response to ischemic injury

BMP-2 induces the expression of chondrocyte-specific genes in bovine synovium- derived progenitor cells cultured in three-dimensional alginate hydrogel

Decellularized vein as a potential scaffold for vascular tissue engineering Patrick J Schaner, MD, Niels D Martin, MD, Thomas N Tulenko, PhD, Irving M.

Undifferentiated mesenchymal stem cells seeded on a vascular prosthesis contribute to the restoration of a physiologic vascular wall Alain Mirza, MD,

Differential effects of orbital and laminar shear stress on endothelial cells Alan Dardik, MD, PhD, Leiling Chen, MD, Jared Frattini, MD, Hidenori Asada,

Sustained orbital shear stress stimulates smooth muscle cell proliferation via the extracellular signal-regulated protein kinase 1/2 pathway Hidenori.

The therapeutic effect of vascular endothelial growth factor gene- or heme oxygenase-1 gene-modified endothelial progenitor cells on neovascularization.

The influence of early-phase remodeling events on the biomechanical properties of engineered vascular tissues Zehra Tosun, Carolina Villegas-Montoya,

Effect of tissue plasminogen activator on vascular smooth muscle cells

Bertrand Poussier, MD, Alfredo C

Angiogenic effects of stromal cell-derived factor-1 (SDF-1/CXCL12) variants in vitro and the in vivo expressions of CXCL12 variants and CXCR4 in human.

Two-wavelength near-infrared fluorescence for the quantitation of drug antiplatelet effects in large animal model systems Yoshitomo Ashitate, MD, Soon.

Tissue engineering applications to vascular bypass graft development: The use of adipose-derived stem cells Paul DiMuzio, MD, Thomas Tulenko, PhD Journal.

Autologous human plasma in stem cell culture and cryopreservation in the creation of a tissue-engineered vascular graft Ping Zhang, PhD, Aleksandra Policha,

Local lentiviral short hairpin RNA silencing of CCR2 inhibits vein graft thickening in hypercholesterolemic apolipoprotein E3-Leiden mice Daniël Eefting,

Bone Marrow-Derived Mesenchymal Stem Cells Expressing Thioredoxin 1 Attenuate Bleomycin-Induced Skin Fibrosis and Oxidative Stress in Scleroderma Miao.

Engineering Patient-Specific Valves Using Stem Cells Generated From Skin Biopsy Specimens David L. Simpson, PhD, Brody Wehman, MD, Yekaterina Galat,

Blood-derived smooth muscle cells as a target for gene delivery

Combination of stromal-derived factor-1α and vascular endothelial growth factor gene- modified endothelial progenitor cells is more effective for ischemic.

Use of Brilliant Blue FCF during vein graft preparation inhibits intimal hyperplasia Michael J. Osgood, MD, Kevin Sexton, MD, Igor Voskresensky, MD, Kyle.

The differential effect of contrast agents on endothelial cell and smooth muscle cell growth in vitro Carol J. Sawmiller, MD, Richard J. Powell, MD,

Foxc2 overexpression enhances benefit of endothelial progenitor cells for inhibiting neointimal formation by promoting CXCR4-dependent homing Dujuan.

Increased connexin43 expression in human saphenous veins in culture is associated with intimal hyperplasia Sébastien Déglise, MD, David Martin, MS, Hervé.

Thomas E. Arnold, MD, Dmitri Gnatenko, PhD, Wadie F. Bahou, MD

Decellularized vein as a potential scaffold for vascular tissue engineering Patrick J Schaner, MD, Niels D Martin, MD, Thomas N Tulenko, PhD, Irving M.

Origin of endothelial cells that line expanded polytetrafluoroethylene vascular grafts sodded with cells from microvascularized fat Stuart K. Williams,

Nicholas P. Ziats, PhD, James M. Anderson, MD, PhD

Sandro Lepidi, MD, Richard D. Kenagy, PhD, Elaine W

Tissue-engineered blood vessel substitute by reconstruction of endothelium using mesenchymal stem cells induced by platelet growth factors Matheus Bertanha,

Daniel J. Wong, MD, Daniel Y. Lu, MD, Clinton D

Murine abdominal aortic aneurysm model by orthotopic allograft transplantation of elastase-treated abdominal aorta Zhenjie Liu, MD, PhD, Qiwei Wang,

Tissue factor pathway inhibitor-2 is induced by fluid shear stress in vascular smooth muscle cells and affects cell proliferation and survival Johan.

Volume 7, Issue 4, Pages (October 2016)

Smooth muscle cells cultured from human saphenous vein exhibit increased proliferation, invasion, and mitogen-activated protein kinase activation in vitro.

Fibronectin promotes VEGF-induced CD34+ cell differentiation into endothelial cells Errol S Wijelath, PhD, Salman Rahman, PhD, Jacqueline Murray, PhD,

An “off the shelf” vascular allograft supports angiogenic growth in three-dimensional tissue engineering Johann M. Zdolsek, MD, Wayne A. Morrison, MB.

Umbilical Cord Blood Derived Endothelial Progenitor Cells for Tissue Engineering of Vascular Grafts Dörthe Schmidt, MD, Christian Breymann, MD, Alberto.

Midkine is expressed by infiltrating macrophages in in-stent restenosis in hypercholesterolemic rabbits Hiroshi Narita, MD, Sen Chen, PhD, Kimihiro Komori,

Leptin receptor is elevated in carotid plaques from neurologically symptomatic patients and positively correlated with augmented macrophage density Jacob.

The proliferative response to platelet-derived growth factor of smooth muscle cells isolated from synthetic vascular grafts in a canine model David J.

Adventitial endothelial implants reduce matrix metalloproteinase-2 expression and increase luminal diameter in porcine arteriovenous grafts Helen M.

The fate of an endothelium layer after preconditioning

Low calcium levels in serum-free media maintain chondrocyte phenotype in monolayer culture and reduce chondrocyte aggregation in suspension culture A.

Navin K. Kapur, MD, Ce Bian, MD, Edward Lin, MD, Clayton B

Bret A. Mettler, MD, Virna L. Sales, MD, Chaz L

Volume 22, Issue 4, Pages (April 2014)

A depleting antibody toward sca-1 mitigates a surge of CD34+/c-kit+ progenitors and reduces vascular restenosis in a murine vascular injury model Bryan.

Tormod S. Westvik, MD, Tamara N

T. Kurth, M. Sc. , E. Hedbom, Ph. D. , N. Shintani, Ph. D. , M

Volume 21, Issue 2, Pages (February 2013)

Retroviral-mediated transduction of endothelial cells with the lac Z gene impairs cellular proliferation in vitro and graft endothelialization in vivo

Aortic wall cell proliferation via basic fibroblast growth factor gene transfer limits progression of experimental abdominal aortic aneurysm Katsuyuki.

Yunge Zhao, MD, PhD, Mark Turlington, BS, Damien J. LaPar, MD, David R

Γ-irradiation modulates vascular smooth muscle cell and extracellular matrix function: implications for neointimal development Joerg Heckenkamp, MD,

The Development and Characterization of an In Vitro Model of Psoriasis

Role of thrombin in endothelial cell monolayer repair in vitro

Intracellular calcium response of endothelial cells exposed to flow in the presence of thrombin or histamine Laura M. Worthen, MS, Matthias U. Nollert,

The Use of Human Sweat Gland–Derived Stem Cells for Enhancing Vascularization during Dermal Regeneration Sandra Danner, Mathias Kremer, Anna Emilia Petschnik,

Jens Hasskarl, Palanivel Velupillai, Karl Münger

Functional analysis of cryopreserved veins

Blockade of monocyte chemoattractant protein-1 by adenoviral gene transfer inhibits experimental vein graft neointimal formation Hideki Tatewaki, MD,

Adult human endothelial cell enzymatic harvesting

Vascular Endothelial Growth Factor Receptor Upregulation in Response to Cell-Based Angiogenic Gene Therapy Terrence M. Yau, MD, MS, Guangming Li, MD,

Panagiotis Kougias, MD, Hong Chai, MD, PhD, Peter H. Lin, MD, Alan B

Presentation transcript:

Autologous human plasma in stem cell culture and cryopreservation in the creation of a tissue-engineered vascular graft Ping Zhang, PhD, Aleksandra Policha, MD, Thomas Tulenko, PhD, Paul DiMuzio, MD Journal of Vascular Surgery Volume 63, Issue 3, Pages 805-814 (March 2016) DOI: 10.1016/j.jvs.2014.10.015 Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 1 Morphology and expansion of human adipose-derived stem cells (ASCs) cultured with autologous human plasma (HP) vs fetal bovine serum (FBS). A, Phase contrast micrographs (20×) of human ASCs (passage 1) grown in medium supplemented with autologous HP (ASC-HP, left) vs FBS (ASC-FBS, right) for 7 days, demonstrating no significant morphologic change. B, Growth curves of human ASCs cultured in medium supplemented with autologous HP vs FBS during 14 days (n = 10). C, Bar graph of doubling times of human ASCs generated from growth curves (n = 10) revealing the trend toward a shorter doubling time for cells grown in HP. Journal of Vascular Surgery 2016 63, 805-814DOI: (10.1016/j.jvs.2014.10.015) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 2 Effect of autologous human plasma (HP) or fetal bovine serum (FBS) on endothelial differentiation of human adipose-derived stem cells (ASCs). A, Expression of mRNA levels of endothelial cell (EC) markers in ASCs. CD31 and von Willebrand factor (vWF) message expression quantified by real-time reverse transcription-polymerase chain reaction (RT-PCR) demonstrates that substitution of the HP resulted in increased expression of these endothelial molecular markers, although not significantly (n = 6; average ± standard error). B, Immunofluorescence micrograph (20×) of ASCs cultured in EGM-2 supplemented with autologous HP vs FBS for 10 days revealing uptake of DiI-labeled acetylated low-density lipoprotein (acLDL) and human lectin under both conditions (red, LDL; green, lectin). ASCs cultured in M199 medium were used as undifferentiated controls. C, Phase contrast photomicrograph (20×) of the differentiated cells subsequently plated onto Matrigel for 12 hours demonstrating the formation of cord-like structures under both culture conditions (representative pictures from two experiments from two different donors). Journal of Vascular Surgery 2016 63, 805-814DOI: (10.1016/j.jvs.2014.10.015) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 3 Creation of a tissue-engineered vascular graft (TEVG) composed of endothelial-differentiated adipose-derived stem cells (ASCs). Laser confocal photomicrographs (20×; CellTracker green) of the luminal surface of vascular grafts (decellularized human saphenous vein) seeded with ASCs and flow conditioned for 5 days (0-9 dyne). A confluent monolayer was achieved with cells cultured in EGM-2 supplemented with either autologous human plasma (HP) or fetal bovine serum (FBS). The micrographs are representative of three grafts created from three different donors. Journal of Vascular Surgery 2016 63, 805-814DOI: (10.1016/j.jvs.2014.10.015) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 4 Proliferation potential and viability of cryopreserved adipose-derived stem cells (ASCs). ASCs differentiated toward endothelial cells (ECs) were cryopreserved in freezing solution (5% dimethyl sulfoxide [DMSO] plus 95% autologous human plasma [HP]) for 1 month. The thawed ASCs were then plated in fresh EGM-2 culture medium. A, Proliferation of HP-ASCs measured by MTT assay before and after cryopreservation reveals preserved proliferation capacity after thawing. Bars show the mean (+standard error of the mean [SEM]) (n = 3). B, Similarly, the proliferation of ASCs cultured in EGM-2 medium supplemented with autologous HP vs fetal bovine serum (FBS) after cryopreservation is preserved. Bars show the mean (+SEM) (n = 3). C, Viability assessment of frozen/thawed ASCs after exposure to different concentrations of DMSO. ASCs cryopreserved in HP or FBS freezing solution containing 5% or 10% DMSO for 2 weeks. Cell viability was determined by the exclusion of trypan blue. Bars show the mean (+SEM) (n = 4). D, Cell survival rate of long-term cryopreserved ASCs was directly measured after thawing by use of the trypan blue assay. The ASCs were stored in HP or FBS freezing solution containing 5% DMSO for short-term (2 weeks) or long-term (>1 year) cryopreservation. Bars show the mean (+SEM) (n = 4). Journal of Vascular Surgery 2016 63, 805-814DOI: (10.1016/j.jvs.2014.10.015) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 5 Expression of endothelial cell (EC) markers and angiogenic potential of cryopreserved adipose-derived stem cells (ASCs). A, Real-time reverse transcription-polymerase chain reaction (RT-PCR) evaluating CD31, von Willebrand factor (vWF), and CD144 message expression in ASCs cultured in medium supplemented with autologous human plasma (HP) vs fetal bovine serum (FBS) before and after cryopreservation revealing preserved expression of these endothelial markers with the exception of the decrease observed in vWF and CD144 in cells cultured in HP. The bars show the mean (+standard error of the mean) (n = 3). B, Representative phase contrast photomicrographs (20×) of ASCs plated on Matrigel for 12 hours before and after cryopreservation revealing preserved ability to form cord-like structures (n = 3). Journal of Vascular Surgery 2016 63, 805-814DOI: (10.1016/j.jvs.2014.10.015) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 6 Cryopreservation of tissue-engineered vascular graft (TEVG). A, Photograph of a TEVG composed of decellularized human greater saphenous vein seeded with adipose-derived stem cells (ASCs) (luminal surface) and flow conditioned with bioreactor for 5 days. B and C, Representative confocal micrographs (10×; CellTracker green) of the luminal surfaces of a TEVG seeded with ASCs cultured in EGM-2 supplemented with autologous human plasma (HP, B) vs fetal bovine serum (FBS, C) and subsequently cryopreserved for 10 days in dimethyl sulfoxide (DMSO)/autologous HP vs FBS freezing solution (left). Comparison with the luminal surface of the same graft before freezing (right) reveals preservation of a confluent luminal surface. Journal of Vascular Surgery 2016 63, 805-814DOI: (10.1016/j.jvs.2014.10.015) Copyright © 2016 Society for Vascular Surgery Terms and Conditions