measuring-respiratory-function-mice-using-unrestrained-whole-body

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

isolation-cryopreservation-culture-human-amnion-epithelial-cells-for

Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications

Human amnion epithelial cells (hAECs) derived from term or pre-term amnion membranes have attracted attention from researchers and clinicians as a ...

Since current treatments for both acute and chronic lung diseases are less than ideal, there has been recent interest in the use of cell-based therapies for inflammatory lung disease. Specifically, human amnion epithelial cells (hAECs) have been shown to reduce bleomycin-induced lung injury and prevent subsequent loss of respiratory function, primarily through modulation of the host immune response. The precise mechanisms of this effect remain unclear. We aimed to investigate the potential of hAECs to mitigate bleomycin-induced lung injury in surfactant protein C deficient (Sftpc(-/-)) mice, which are highly susceptible to pulmonary injury as a result of impairment of macrophage function. Primary hAECs were administered to wild type (Sftpc(+/+)) and Sftpc(-/-) mice 24 h after exposure to bleomycin. Compared to Sftpc(+/+) mice receiving bleomycin alone, Sftpc(+/+) mice administered hAECs 24 hours after bleomycin exposure had decreased expression of proinflammatory genes, decreased macrophage and neutrophil infiltration, fibrosis, collagen content and Î±-smooth muscle actin as well as a significant improvement in lung function. Compared to Sftpc(-/-) mice given bleomycin alone, Sftpc(-/-) mice administered hAECs 24 hours after bleomycin, did not have a decrease in inflammatory gene expression or a reduction in macrophage pulmonary infiltration. Subsequently, Sftpc(-/-) mice did not show any decrease in pulmonary fibrosis or improvement of lung function after hAEC administration. The ability of hAECs to mitigate bleomycininduced lung injury is abolished in Sftpc(-/-) mice, suggesting that hAECs require normal host macrophage function to exert their reparative effects.

HUMAN AMNION EPITHELIAL CELLS DO NOT ABROGATE PULMONARY FIBROSIS IN MICE WITH IMPAIRED MACROPHAGE FUNCTION.

Aktuelle Behandlungen f&uuml;r beide akute und chronische Lungenerkrankungen sind weniger als ideal, seit den letzten Interesse am Einsatz von zellbasierte Therapien f&uuml;r entz&uuml;ndliche Lungenerkrankung. Insbesondere wurden menschliche Amnion Epithelzellen (hAECs) gezeigt, zur Verringerung Bleomycin-induzierte Lungeninsuffizienz und nachfolgende Verlust von Atmung, vor allem durch Modulation der Immunantwort Host zu verhindern. Die genauen Mechanismen f&uuml;r diesen Effekt sind noch unklar. Wir wollten untersuchen das Potenzial der hAECs zur Minderung von Bleomycin-induzierte Respiratory Distress in Tensid Protein C-Mangel (Sftpc(-/-)) M&auml;use, die sehr anf&auml;llig f&uuml;r pulmonale Sch&auml;digung infolge Beeintr&auml;chtigung der Funktion der Makrophagen sind. Prim&auml;re hAECs wurden Wildtyp (Sftpc(+/+)) und Sftpc(-/-) M&auml;use 24 h nach Exposition mit Bleomycin verabreicht. Im Vergleich zu Sftpc(+/+) M&auml;use empfangen von Bleomycin allein, verabreicht Sftpc(+/+) M&auml;usen hAECs 24 Stunden nach Bleomycin Exposition Ausdruck der Proinflammatory Gene, verringerte Makrophagen und neutrophile Infiltration, Fibrose, Kollagen Inhalt und &alpha;-glatten Muskel Aktin sowie eine deutliche Verbesserung der Lungenfunktion zur&uuml;ckgegangen. Im Vergleich zu Sftpc(-/-) M&auml;use Bleomycin allein gegeben, Sftpc(-/-) M&auml;usen verabreicht hAECs 24 Stunden nach Bleomycin, keine Abnahme der entz&uuml;ndlichen Genexpression oder eine Verringerung der Makrophagen pulmonale Infiltration. Anschlie&szlig;end zeigte die Sftpc(-/-) M&auml;use kein R&uuml;ckgang der Lungenfibrose oder Verbesserung der Lungenfunktion nach hAEC Verabreichung. Die F&auml;higkeit von hAECs, Bleomycininduced Respiratory Distress Syndrome zu mildern ist abgeschafft, in Sftpc(-/-) M&auml;use, darauf hindeutet, dass hAECs normalen Host Makrophage-Funktion ihrer Reparativen Wirkungen aus&uuml;ben werden.

MENSCHLICHE AMNION EPITHELZELLEN NICHT AUFHEBT, LUNGENFIBROSE IN M&Auml;USE MIT GEST&Ouml;RTER MAKROPHAGE-FUNKTION.

In the United States alone, there are approximately 500,000 burn injuries that require medical treatment every year. Limitations of current treatments necessitate the development of new methods that can be applied quicker, result in faster wound regeneration, and yield skin that is cosmetically similar to undamaged skin. The development of new hydrogel biomaterials and bioprinting deposition technologies has provided a platform to address this need. Herein we evaluated characteristics of twelve hydrogels to determine their suitability for bioprinting applications. We chose hydrogels that are either commercially available, or are commonly used for research purposes. We evaluated specific hydrogel properties relevant to bioprinting applications, specifically; gelation time, swelling or contraction, stability, biocompatibility and printability. Further, we described regulatory, commercial and financial aspects of each of the hydrogels. While many of the hydrogels screened may exhibit characteristics suitable for other applications, UV-crosslinked Extracel, a hyaluronic acid-based hydrogel, had many of the desired properties for our bioprinting application. Taken together with commercial availability, shelf life, potential for regulatory approval and ease of use, these materials hold the potential to be further developed into fast and effective wound healing treatments.

Evaluation of hydrogels for bio-printing applications.

Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure for cystic fibrosis. With this in mind, we investigated the ability of human amnion epithelial cells (hAECs) to express functional CFTR. We found that hAECs formed 3-dimensional structures and expressed the CFTR gene and protein after culture in Small Airway Growth Medium (SAGM). We also observed a polarized CFTR distribution on the membrane of hAECs cultured in SAGM, similar to that observed in polarized airway cells in vivo. Further, hAECs induced to express CFTR possessed functional iodide/chloride (I(-/)Cl(-)) ion channels that were inhibited by the CFTR-inhibitor CFTR-172, indicating the presence of functional CFTR ion channels. These data suggest that hAECs may be a promising source for the development of a cellular therapy for cystic fibrosis.

Human amnion epithelial cells induced to express functional cystic fibrosis transmembrane conductance regulator.

Zystische Fibrose, autosomal-rezessiv eine Mutation in einem Gen Kodierung der Cystic Fibrosis transmembrane Conductance Regulator (CFTR), bleibt eine f&uuml;hrende Ursache von Kindheit respiratorische Morbidit&auml;t und Mortalit&auml;t. Die Atemwege Folgen der zystischen Fibrose sind die Generation von dicken, z&auml;hen Schleim, die Lunge Zollabwicklung, pr&auml;disponierenden einzelnen wiederholte und anhaltende Infektionen, progressives Lungensch&auml;den und verk&uuml;rzte Lebensdauer beeintr&auml;chtigt. Derzeit gibt es keine Heilung f&uuml;r Mukoviszidose. In diesem Sinne untersuchten wir die F&auml;higkeit der menschlichen Amnion Epithelzellen (hAECs), funktionelle CFTR auszudr&uuml;cken. Wir fanden, dass hAECs 3-dimensionale Strukturen gebildet und die CFTR-Gen und Protein nach Kultur in kleinen Atemwege Wachstum Medium (SAGM) ausgedr&uuml;ckt. Wir beobachten auch eine polarisierte CFTR-Verteilung auf der Membran des hAECs kultiviert in SAGM, &auml;hnlich, die im polarisierten Airway-Zellen in vivo beobachtet. Dar&uuml;ber hinaus induzierte hAECs CFTR Ausdr&uuml;cken besa&szlig; funktionale Jodid/Chlorid (I(-/)Cl(-)) Ionenkan&auml;le, die durch die CFTR-Inhibitor CFTR-172, das Vorhandensein von funktionellen CFTR-Ionenkan&auml;len behindert wurden. Diese Daten legen nahe, dass hAECs m&ouml;glicherweise eine vielversprechende Quelle f&uuml;r die Entwicklung einer zellul&auml;ren Therapie f&uuml;r Mukoviszidose.

Menschliche Amnion Epithelzellen induziert funktionale zystische Fibrose Transmembran Conductance Regulator zum Ausdruck zu bringen.

Background: The immunomodulatory and immunosuppressive capacity of human mesenchymal stem cells (hMSC) is well recognized, but efficacies of hMSC in immunocompetent and immunocompromised animals have never been directly compared. Objectives: We aimed to compare the efficacy of hMSC in preventing bleomycin-induced lung injury in immunocompromised SCID and immunocompetent C57Bl/6 mice. Methods: SCID and C57Bl/6 mice were subjected to a single bolus intranasal instillation of bleomycin to induce lung injury. One million hMSC were administered intravenously 24 h following the induction of bleomycin lung injury. Results: hMSC xenotransplantation into SCID mice resulted in transient improvements in lung weight and tidal volume and to persistent improvement in inspiratory duty cycle, inspiratory flow rate and inspiration/expiration ratio. We did not observed protective effects in C57Bl/6 mice. This correlated with histological changes, where hMSC administration reduced Ashcroft scores, collagen deposition and inflammatory influx in the lungs of SCID mice, but not in those of C57Bl/6 mice. Conclusion: The application of hMSC for the treatment of acute and chronic lung injury is significantly affected by the immune status of the recipient. Lack of hMSC-mediated repair observed in C57Bl/6 mice was likely to be due to limitations of their immune privilege and differential priming of hMSC in immunocompetent versus immunocompromised hosts.

Human Mesenchymal Stem Cells Reduce Lung Injury in Immunocompromised Mice but Not in Immunocompetent Mice.

Hintergrund: Die immunmodulatorische und immunsuppressive Kapazit&auml;t menschlicher mesenchymale Stammzellen (hMSC) wurde auch erkannt, aber Leistungsnachteile der hMSC bei immunkompetenten und immungeschw&auml;chten Tieren nie direkt verglichen wurden. Ziele: Wir vergleichen die Wirksamkeit von hMSC Bleomycin-induzierte Respiratory Distress Syndrome bei immungeschw&auml;chten SCID und immunkompetenten C57Bl/6 M&auml;usen zu verhindern soll. Methoden: SCID und C57Bl/6 M&auml;usen wurden ein einzelnes Bolus intranasale Instillation von Bleomycin Lungeninsuffizienz veranlassen unterworfen. 1 Million hMSC wurden intraven&ouml;s verabreicht 24 h nach die Induktion von Bleomycin Respiratory Distress Syndrome. Ergebnisse: hMSC Xenotransplantation in SCID M&auml;use f&uuml;hrte zu vor&uuml;bergehenden Verbesserungen Lunge Gewichts-und Tidalvolumen und anhaltende Verbesserung inspiratorischen Einschaltdauer, inspiratorischen Flow Rate und Inspiration/Ablauf-Verh&auml;ltnis. Wir haben nicht beobachteten sch&uuml;tzende Wirkung bei C57Bl/6 M&auml;usen. Dies korreliert mit den histologischen Ver&auml;nderungen, wo hMSC Verwaltung Ashcroft reduziert, Livescores, Ablagerung von Kollagen und entz&uuml;ndliche Zustrom in die Lungen der SCID-M&auml;use, aber nicht in denen der C57Bl/6 M&auml;usen. Fazit: Die Anwendung der hMSC zur Behandlung von akuten und chronischen Lungeninsuffizienz wird von den Immunstatus des Empf&auml;ngers erheblich beeinflusst. Mangelnde hMSC-vermittelte Reparatur bei C57Bl/6 M&auml;usen beobachtet wurde wahrscheinlich wegen Einschr&auml;nkungen der ihr immun-Privileg und differenzielle Grundierung des hMSC in immunkompetenten gegen&uuml;ber immungeschw&auml;chten Gastgeber.

Humaner mesenchymaler Stammzellen verringern Respiratory Distress Syndrome bei immungeschw&auml;chten M&auml;usen, aber nicht bei immunkompetenten M&auml;usen.

Gestational tissue such as the placenta, placental membranes, and amniotic fluid are usually discarded following birth. Recently, researchers have identified gestational tissue as an untapped source of stem cells that are highly multipotent and possess potent immunosuppressive properties. Placental mesenchymal stem cells (MSCs), human amnion epithelial cells (hAECs), and amniotic fluid-derived stem cells (AFSCs) have been shown to differentiate into various cell types including adipogenic, osteogenic, myogenic, endothelial, pulmonary, neurogenic, hepatogenic, cardiac, and pancreatic lineages. Their immunomodulatory properties suggest that gestational stem cells may have an important application in the treatment of various inflammatory diseases such as graft versus host and autoimmune diseases. In clinical and preclinical studies, gestational stem cells have shown efficacy in the treatment of Crohn disease, lung disease, diabetes, repair of bone defects, heart disease, kidney disease, neural degeneration, and blood disorders. Stem cells derived from the placenta, placental membranes, and amniotic fluid are a valuable resource for the field of regenerative medicine.

Amniotic fluid and placental membranes: unexpected sources of highly multipotent cells.

Currently there is no cure for cystic fibrosis (CF). Treatments are focused on addressing the disease symptoms, with varying degrees of success. Regenerative medicine holds the promise of regenerating dysfunctional or damaged tissues and to enhance the body's own endogenous repair mechanisms. The discovery of endogenous and exogenous stem cells has provided valuable tools for development of novel treatments for CF. The ability of stem cells to differentiate into functional pulmonary cells, modulate inflammatory responses and contribute to pulmonary function has provided researchers with multiple approaches to develop effective treatment strategies. Several approaches show promise to produce viable therapeutic treatments to treat the underlying cause of CF, reduce the symptoms and mitigate long-term damage, and generate functional replacement organs for end-stage transplantation. This review provides an overview of the rapidly progressing field of cell therapy for CF, focusing on the various cell types utilized and current strategies that show promise to improve life expectancy and quality of life for CF patients. Copyright © 2013 John Wiley & Sons, Ltd.

Cell therapy for cystic fibrosis.

Over the last few decades, both synthetic and natural materials have been utilized to develop bladder substitutes. Most attempts have not been successful because of mechanical, structural, functional, or biocompatibility problems. Bladder acellular matrix (BAM) is obtained by removing cellular components from donor bladders, leaving a tissue matrix consisting of collagen, elastin, fibronectin, glycosaminoglycans, proteoglycans, and growth factors. Multiple BAM-based studies now suggest that tissue engineering techniques may provide efficacious alternatives to current methods of bladder augmentation. Efforts to optimize BAM-based scaffolds are ongoing and would be greatly assisted by feasible means of improving scaffold properties and interaction with cells and tissues. Future applications of BAM will likely include cell-seeded grafts with the eventual hope of producing "off the shelf" replacement materials for bladder augmentation.

Bladder acellular matrix and its application in bladder augmentation.

Tracheal reconstruction using a stem cell-based engineered trachea has recently shown promise. Our goal is to achieve a single-stage stem cell-based tracheal replacement.

Tracheal reconstruction in a canine model.

Repair and reconstruction of damaged tissues and organs has been a major issue in the medical field. Regenerative medicine and tissue engineering, as rapid evolving technologies, may offer alternative treatments and hope for patients with serious defects and end-stage diseases. Most urologic diseases could benefit from the development of regenerative medicine and tissue engineering. This article discusses the role of cells and materials in regenerative medicine, as well as the status of current role of regenerative medicine for the generation of specific urologic organs.

Regenerative medicine in urology.

Additive manufacturing, otherwise known as three-dimensional (3D) printing, is driving major innovations in many areas, such as engineering, manufacturing, art, education and medicine. Recent advances have enabled 3D printing of biocompatible materials, cells and supporting components into complex 3D functional living tissues. 3D bioprinting is being applied to regenerative medicine to address the need for tissues and organs suitable for transplantation. Compared with non-biological printing, 3D bioprinting involves additional complexities, such as the choice of materials, cell types, growth and differentiation factors, and technical challenges related to the sensitivities of living cells and the construction of tissues. Addressing these complexities requires the integration of technologies from the fields of engineering, biomaterials science, cell biology, physics and medicine. 3D bioprinting has already been used for the generation and transplantation of several tissues, including multilayered skin, bone, vascular grafts, tracheal splints, heart tissue and cartilaginous structures. Other applications include developing high-throughput 3D-bioprinted tissue models for research, drug discovery and toxicology.

ArticleTotal : 2	Year
Publication title Cited by 69	2014
Publication title Cited by 29	2014

Article	Year
HUMAN AMNION EPITHELIAL CELLS DO NOT ABROGATE PULMONARY FIBROSIS IN MICE WITH IMPAIRED MACROPHAGE FUNCTION. Cell transplantation\| PubMed ID: 22507554	2012
Evaluation of hydrogels for bio-printing applications. Journal of biomedical materials research. Part A\| PubMed ID: 22941807	2013
Human amnion epithelial cells induced to express functional cystic fibrosis transmembrane conductance regulator. PloS one\| PubMed ID: 23029546	2012
Human Mesenchymal Stem Cells Reduce Lung Injury in Immunocompromised Mice but Not in Immunocompetent Mice. Respiration; international review of thoracic diseases\| PubMed ID: 23207668	2012
Amniotic fluid and placental membranes: unexpected sources of highly multipotent cells. Seminars in reproductive medicine\| PubMed ID: 23329638	2013
Cell therapy for cystic fibrosis. Journal of tissue engineering and regenerative medicine\| PubMed ID: 23894126	2013
Bladder acellular matrix and its application in bladder augmentation. Tissue engineering. Part B, Reviews\| PubMed ID: 23895225	2014
Tracheal reconstruction in a canine model. Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery\| PubMed ID: 24367052	2014
Regenerative medicine in urology. Seminars in pediatric surgery\| PubMed ID: 24994523	2014
3D bioprinting of tissues and organs. Nature biotechnology\| PubMed ID: 25093879	2014

Sean V. Murphy

Biography

JoVE Journal Publications

Other Publications