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Thoraxchirurgische Forschung


 

Projekte

Establishment of primary pleural mesothelium with 3D organotypic coculture

Background/Purpose

Pleural mesothelial cells (PMCs) lead the pleural immune response and serve as progenitor cells after pleural or lung damage. They therefore play a key role in pleural diseases such as bacterial infections, malignant pleural effusion, pleural carcinosis or pleural mesothelioma. All these diseases prove a significant healthcare burden, however, its scientific consideration is quite low. The aim of the present study was to develop a three-dimensional (3D) in vitro organotypic model for investigation of pathological conditions of the pleural mesothelium. 3D organotypic models are a promising approach to gain an in vivo like understanding of molecular disease development. While these models are well established for a variety of organs, such as lung, colon, intestine, liver, pancreas, esophagus, prostate, or omentum majus, a 3D organoid model of the pleura is still missing. 

Methods

To construct a 3D organotypic model, primary human PMCs and fibroblasts were isolated from human pleura biopsies. Purification of primary human PMCs and fibroblasts was verified by immunofluorescence staining. The 3D collagen gel culture was assembled by plating of human pleural fibroblasts inside the gel, followed by seeding of PMCs on the gel to construct the normal pleura. 

Results 

Isolated human PMCs showed cobblestone appearance and expressed mesenchymal characteristics: αSMA, vimentin but not prolylhydroxylase 1 (PHD1). The extracted fibroblasts maintained their spindle cell appearance and were positive for PHD1 (fibroblast marker). Grown on top of matrix-embedded fibroblasts, the primary human PMCs establish a monolayer and have direct contact with the underlying fibroblasts. Forty-eight hours after attachment, PMCs had cobblestone appearance and intercellular junctions were present between the mesothelial cells as shown by immunostaining for ZO-1. The structural and functional phenotype of the PMCs in our 3D organotypic culture was preserved over six days of culture, as evidenced by the expression of mesenchymal (vimentin, α-SMA, ZO-1) and proliferation marker (Ki67). Conclusions

The presented 3D organotypic model of pleura functions as a robust assay for pleural research serving as a precise reproduction of the in vivo morphology and microenvironment and presents a novel tool for development of preventive and therapeutic enhancement of various pleural diseases.​


 

Untersuchungen zur Pathogenese apikaler bullöser Veränderungen in den Lungenspitzen im Segment 1 & Segment 6

Das Auftreten des primären Spontanpneumothorax trifft vorwiegend jugendliche Patienten. Klinisch relevant ist dabei die hohe Rezidivrate die damit einhergeht. Zum jetzigen Zeitpunkt ist die Pathogenese dieser apikal bullösen Veränderungen der Lungenspitzen im Bereich Segment 1 und Segment 6 des jeweiligen Lungenflügels aufgrund der klinischen Situation von großem Interesse. Bei den hier geplanten Untersuchungen ist von Bedeutung, die Mechanismen der bullösen Veränderungen zu charakterisieren, die Lokalisation und die molekularen Ursachen im Defekt der Lungenstruktur zu identifizieren. Vorwiegend ist von Interesse die Untersuchung der idiopathischen juvenilen Pneumothoraces. Dabei werden in der Studie Gewebe von Lunge und Pleura von Patienten mit entsprechender Indikation für chirurgische Intenvention im Labor ex vivo histologisch und molekularbiologisch analysiert und in vitro kultiviert. Fokussiert wird bei den Analysen vor allem auf mögliche Veränderungen der extrazellulären Matrix, Veränderungen im Regenerationsverhalten, auf Veränderungen durch epitheliale-mesenchymale Transition der Epithelzellen und Apoptoseverhalten. Zusätzlich werden in vitro Studien mit der alveolarepithelialen Lungenadenokarzinomzelllinie A549 durchgeführt, um den Einfluss von mesenchymalen Stromazellen als mögliche zelltherapeutische Intervention zu erfassen.

Team

Leiter: Prof. Dr. med. Matthias Steinert

 
Dr. med. Dr. rer. med. Isabella Metelmann, MA

 
Dr. med. Sebastian Krämer

 
Dr. rer. nat. Olga Kurow

cand. med. Melinda Yorgun

 
Dr. rer. nat. Peggy Stock

 

 

Aktuelles

​ ​​DGT

31. Jahrestagung der Deutschen Gesellschaft für Thoraxchirurgie vom 12.-14. September 2022 in Essen​


 

 

 
CTRMS 

 

Dr. Peggy Stock aus der AG Angewandte Molekulare Hepatologie um Prof. Bruno Christ wurde für weitere 4 Jahre im Beirat der Cell Transplantation and Regenerative Medicine Society (CTRMS) bestätigt. Die Wissenschaftlerin aus der Klinik und Poliklinik für Viszeral-, Transplantations-, Thorax- und Gefäßchirurgie übernimmt in der internationalen Gesellschaft, welche unter dem Dach der TTS (The Transplantation Society) agiert, bis 2025 neben den Aufgaben eines Beiratsmitglieds auch die Leitung des Young Investigators Committee der CTRMS.

Zusammen mit der Medizinstudentin Frau Melinda Yorgun, welche im Bereich Thoraxchirurgie der VTTG unter der Leitung von Prof. Matthias Steinert promoviert, und sich ebenfalls als Trainee Member der CTRMS engagiert, unterstützen sie aktiv die Interaktionen zwischen den jüngeren Wissenschaftlern und etablierten Mitgliedern auf dem Gebieten der Zelltherapie und der regenerativen Medizin.​



 

Publikationen

Team

Dr. Dr. Isabella B. Metelmann

Metelmann IB, Seyfarth HJ, Schierle K, Kraemer S. Utility of immunohistochemical staining for the diagnosis of Extra-adrenal mediastinal paraganglioma. Respir Med Case Rep. 2020 Nov 13;31:101278. doi: 10.1016/j.rmcr.2020.101278. PMID: 33294355; PMCID: PMC7683335.  
 
Metelmann IB, Oltmans A, Kraemer S. Isolated open bronchus fracture after open die forging. Trauma Case Rep. 2021 Mar 17;32:100460. doi: 10.1016/j.tcr.2021.100460. PMID: 33816743; PMCID: PMC8010854. 
 
Broschewitz J, Metelmann I, Steinert M, Krämer S. Thoraxchirurgie ohne Thoraxdrainage – aktuelle Situation in Deutschland [Thoracic Surgery Without Chest Tube: The Current Situation in Germany]. Zentralbl Chir. 2021 Jul 5. German. doi: 10.1055/a-1502-8210. Epub ahead of print. PMID: 34225380.
 
Metelmann I, Broschewitz J, Pietsch UC, Huschak G, Eichfeld U, Bercker S, Kraemer S. Procedural times in early non-intubated VATS program - a propensity score analysis. BMC Anesthesiol. 2021 Feb 11;21(1):44. doi: 10.1186/s12871-021-01270-4. PMID: 33573604; PMCID: PMC7877094.  

 

Dr. Olga Kurow

Präsentation anlässlich der Chirurgischen Forschungstage 2021​


cft2021.png
 


 
Establishment of primary pleural mesothelium with 3D organotypic coculture 

O. Kurow1 , M. Steinert1 , S. Langer2 , S. Kraemer1 , I. Metelmann1 
1 Universitätsklinikum Leipzig, Klinik und Poliklinik für Viszeral-, Transplantations-, Thorax- und Gefäßchirurgie, Leipzig, Germany 
2 Universitätsklinikum Leipzig, Klinik und Poliklinik für Orthopädie, Unfallchirurgie und Plastische Chirurgie, Leipzig, Germany 

 

Background/Purpose 

Pleural mesothelial cells (PMCs) lead the pleural immune response and serve as progenitor cells after pleural or lung damage. They therefore play a key role in pleural diseases such as bacterial infections, malignant pleural effusion, pleural carcinosis or pleural mesothelioma. All these diseases prove a significant healthcare burden, however, its scientific consideration is quite low. The aim of the present study was to develop a three-dimensional (3D) in vitro organotypic model for investigation of pathological conditions of the pleural mesothelium. 3D organotypic models are a promising approach to gain an in vivo like understanding of molecular disease development. While these models are well established for a variety of organs, such as lung, colon, intestine, liver, pancreas, esophagus, prostate, or omentum majus, a 3D organoid model of the pleura is still missing. 

Methods 

To construct a 3D organotypic model, primary human PMCs and fibroblasts were isolated from human pleura biopsies. Purification of primary human PMCs and fibroblasts was verified by immunofluorescence staining. The 3D collagen gel culture was assembled by plating of human pleural fibroblasts inside the gel, followed by seeding of PMCs on the gel to construct the normal pleura. 

Results 

Isolated human PMCs showed cobblestone appearance and expressed mesenchymal characteristics: αSMA, vimentin but not prolylhydroxylase 1 (PHD1). The extracted fibroblasts maintained their spindle cell appearance and were positive for PHD1 (fibroblast marker). Grown on top of matrix-embedded fibroblasts, the primary human PMCs establish a monolayer and have direct contact with the underlying fibroblasts. Forty-eight hours after attachment, PMCs had cobblestone appearance and intercellular junctions were present between the mesothelial cells as shown by immunostaining for ZO-1. The structural and functional phenotype of the PMCs in our 3D organotypic culture was preserved over six days of culture, as evidenced by the expression of mesenchymal (vimentin, α-SMA, ZO-1) and proliferation marker (Ki67). Conclusions 

​The presented 3D organotypic model of pleura functions as a robust assay for pleural research serving as a precise reproduction of the in vivo morphology and microenvironment and presents a novel tool for development of preventive and therapeutic enhancement of various pleural diseases.

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