Name: a 3d human lung tissue model for functional studies on mycobacterium tuberculosis infection
Uploaded: 2015-10-05T16:00:00.000+00:00
Duration: 10 min 10 s
Description: Watch this Scientific Journal Video about A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection at JoVE.com

The authors acknowledge the Microscopy core facility at the Faculty of Health Sciences, Link&#246;ping University for providing access to advanced imaging systems; Karl-Eric Magnusson (Emeritus Scientist) at the Dept. of Clinical and Experimental Medicine, Link&#246;ping University for providing access to Imaris 3D/4D image processing software (Bitplane, Switzerland); and S. Braian for his help with the lung model cartoon. This work was supported by funds from the Swedish Research Council (Alternatives to animal research, 2012-1951) and Swedish Research Council (2012-3349) to M.L. and Swedish Foundation for Strategic Research to S.B. S.B. receive grants from the Karolinska Institutet, Swedish Research Council, the Swedish International Development Cooperation Agency (Sida) and the Swedish Civil Contingencies Agency (MSB), and the Swedish Heart and Lung Foundation (HLF). M.S. received grants from the Karolinska Institutet and Stockholm County Council.

<ol>
	<li>Sakamoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathology+of+Mycobacterium+tuberculosis+infection.">The pathology of Mycobacterium tuberculosis infection.</a> Veterinary pathology. 49, 423-439 (2012).</li><li>Saunders, B. M., Britton, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+and+death+in+the+granuloma:+immunopathology+of+tuberculosis.">Life and death in the granuloma: immunopathology of tuberculosis.</a> Immunol Cell Biol. 85, 103-111 (2007).</li><li>Kaufmann, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+issues+in+tuberculosis.+Ann+Rheum+Dis.">New issues in tuberculosis. Ann Rheum Dis.</a> Ann Rheum Dis. 63, ii50-ii56 (2004).</li><li>Morrison, J., Pai, M., Hopewell, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tuberculosis+and+latent+tuberculosis+infection+in+close+contacts+of+people+with+pulmonary+tuberculosis+in+low-income+and+middle-income+countries:+a+systematic+review+and+meta-analysis.">Tuberculosis and latent tuberculosis infection in close contacts of people with pulmonary tuberculosis in low-income and middle-income countries: a systematic review and meta-analysis.</a> Lancet Infect Dis. 8, 359-368 (2008).</li><li>Barry 3rd, C. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+spectrum+of+latent+tuberculosis:+rethinking+the+biology+and+intervention+strategies.">The spectrum of latent tuberculosis: rethinking the biology and intervention strategies.</a> Nat Rev Microbiol. 7, 845-855 (2009).</li><li>Puissegur, M. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+dual+model+of+mycobacterial+granulomas+to+investigate+the+molecular+interactions+between+mycobacteria+and+human+host+cells.">An in vitro dual model of mycobacterial granulomas to investigate the molecular interactions between mycobacteria and human host cells.</a> Cell Microbiol. 6, 423-433 (2004).</li><li>Kapoor, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+Granuloma+In+Vitro+Model,+for+TB+Dormancy+and+Resuscitation.">Human Granuloma In Vitro Model, for TB Dormancy and Resuscitation.</a> PLoS One. 8, e53657 (2013).</li><li>Nguyen Hoang, ., T, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendritic+cell+functional+properties+in+a+three-dimensional+tissue+model+of+human+lung+mucosa.">Dendritic cell functional properties in a three-dimensional tissue model of human lung mucosa.</a> Am J Physiol Lung Cell Mol Physiol. 302, L226-L237 (2012).</li><li>Parasa, V. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+Mycobacterium+tuberculosis+early+granuloma+formation+in+experimental+human+lung+tissue.">Modeling Mycobacterium tuberculosis early granuloma formation in experimental human lung tissue.</a> Dis Model Mech. 7, 281-288 (2014).</li><li>Cozens, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CFTR+expression+and+chloride+secretion+in+polarized+immortal+human+bronchial+epithelial+cells.">CFTR expression and chloride secretion in polarized immortal human bronchial epithelial cells.</a> Am J Respir Cell Mol Biol. 10, 38-47 (1994).</li><li>Brighenti, S., Andersson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+immune+responses+in+human+tuberculosis:+learning+from+the+site+of+infection.">Local immune responses in human tuberculosis: learning from the site of infection.</a> J Infect Dis. 205, S316-S324 (2012).</li><li>Davis, J. M., Ramakrishnan, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+granuloma+in+expansion+and+dissemination+of+early+tuberculous+infection.">The role of the granuloma in expansion and dissemination of early tuberculous infection.</a> Cell. 136, 37-49 (2009).</li><li>Gupta, U. D., Katoch, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+tuberculosis.">Animal models of tuberculosis.</a> Tuberculosis (Edinb). 85, 277-293 (2005).</li><li>Kashino, S. S., Napolitano, D. R., Skobe, Z., Campos-Neto, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guinea+pig+model+of+Mycobacterium+tuberculosis+latent/dormant+infection.">Guinea pig model of Mycobacterium tuberculosis latent/dormant infection.</a> Microbes Infect. 10, 1469-1476 (2008).</li><li>Singhal, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+tuberculosis+in+the+Wistar+rat:+a+model+for+protective+immunity+and+control+of+infection.">Experimental tuberculosis in the Wistar rat: a model for protective immunity and control of infection.</a> PLoS One. 6, e18632 (2011).</li><li>Subbian, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phosphodiesterase-4+inhibition+alters+gene+expression+and+improves+isoniazid-mediated+clearance+of+Mycobacterium+tuberculosis+in+rabbit+lungs.">Phosphodiesterase-4 inhibition alters gene expression and improves isoniazid-mediated clearance of Mycobacterium tuberculosis in rabbit lungs.</a> PLoS Pathog. 7, e1002262 (2011).</li><li>Lin, P. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+necrosis+factor+neutralization+results+in+disseminated+disease+in+acute+and+latent+Mycobacterium+tuberculosis+infection+with+normal+granuloma+structure+in+a+cynomolgus+macaque+model.">Tumor necrosis factor neutralization results in disseminated disease in acute and latent Mycobacterium tuberculosis infection with normal granuloma structure in a cynomolgus macaque model.</a> Arthritis Rheum. 62, 340-350 (2010).</li><li>Rahman, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Compartmentalization+of+immune+responses+in+human+tuberculosis:+few+CD8++effector+T+cells+but+elevated+levels+of+FoxP3++regulatory+t+cells+in+the+granulomatous+lesions.">Compartmentalization of immune responses in human tuberculosis: few CD8+ effector T cells but elevated levels of FoxP3+ regulatory t cells in the granulomatous lesions.</a> Am J Pathol. 174, 2211-2224 (2009).</li></ol>

The authors declare no competing financial interests.

The ability to recruit and form organized cell clusters at the site of infection is the hallmark of human TB 11. These dynamic structures known as tubercle granulomas primarily consist of immune cells (macrophages, monocytes, T-cells and B-cells) and multi-nucleated giant cells surrounding M. tuberculosis. The role of the granuloma has long been considered to wall off the infection, preventing local spread of bacteria. However, more recent studies show that granuloma formation is critical for early bacterial survival, growth and dissemination 12. A strategy of new studies is to identify molecules or pathways that could efficiently be targeted to inhibit the cellular migration in granuloma formation and/or TB dissemination. 
A caveat for novel studies on TB is the lack of models that recapitulate human TB. The most widely used experimental animals do not form true granuloma upon M. tuberculosis infection, and are therefore not appropriate choices for studies of TB 13-16. Non-human primates have the closest resemblance to human TB 17, but are not the preferred choice owing to high operational costs and ethical issues. Human TB is a complex immunological process and is difficult to model in vitro. Cell cultures of monolayers or co-cultures lack the 3D environment and tissue responses. Therefore, we have developed an innovative lung tissue model based on human primary immune cells and human lung-specific cell lines 8,9. The model displays characteristic features of human lung tissue, including epithelia with evenly integrated macrophages, formation of extracellular matrix, stratified epithelia and mucus secretion 9. 
The 3D human lung tissue model has several benefits over the in vitro single or co-cultures seeded on tissue culture plates or transwell inserts. First, the human lung-specific cells (fibroblasts and epithelial cells) are not commonly included in the in vitro single or co-cultures. Second, the immune cells and lung-specific cells are embedded in a 3D physiological context (collagen rich extra-cellular matrix products). The response of cells to a stimulus/infection and the migratory behaviour of cells, for instance formation of a granuloma, differ significantly between a 2D and 3D environment. Furthermore, the described method enables the 3D visualization and robust 3D quantitative analysis that provides pivotal information on spatial distribution and intricate cellular interactions. 
Experimental infection in the model tissue with M. tuberculosis resulted in clustering of macrophages at the site of infection, reminiscent of early TB granuloma (Figure 2 and 3). We have recently demonstrated that mutant strains defective in the ability to secrete the virulence factor ESAT-6 or Mycobacterium bovis BCG that lacks ESAT-6 did not induce the clustering of monocytes (no early granuloma), in contrast to the virulent M. tuberculosis 9. These data are consistent with the observations made from Mycobacterium marinum-infected zebrafish embryos, whose transparency allows for elegant live imaging of granuloma formation 12. As there is no gold-standard model for TB, we took advantage of the surgically resected tissue biopsies from TB patients for validation of the method 9. Our in vitro tissue model shares several characteristics with the lung and lymph node biopsies from TB patients, including the aggregation of macrophages in granuloma, the presence of both intra- and extracellular bacteria 18 and induction of necrosis 11. 
Although the described model has physiological relevance to human TB and has several advantages over other in vitro models, it has some limitations. For instance, out of more than 20 collagen proteins identified in humans, only type I is included to the model to mimic the extra-cellular matrix. However, type I collagen is a complex mixture of extra-cellular matrix products and is the most abundant collagen in the human body. Further, we have demonstrated the presence of collagen IV and several extra-cellular matrix proteins such as tropoelastin, vimentin and laminin, which are produced by the epithelial cells and fibroblasts in the tissue model, indicating the synthesis of new collagen 8. Presently, the lung tissue model only has monocytes and macrophages, besides lung-specific cells. It lacks neutrophils and lymphocytes that are also known to be present in the granuloma. Remarkably the model is not limited to the introduction of additional immune cells and is of interest to explore how they contribute to the complex cellular interactions in human TB. Implantation of primary alveolar macrophages, skin-specific cells and lung carcinoma cells has already been tested in the model. Since our objective was to use a model that closely resembles human TB, introduction of mouse cells have not been attempted.
In summary, the lung tissue model has implications for both basic mechanistic and applied studies. Potential applications of the lung model include the study of innate immunity, investigating mechanistic aspects of host defences such as phagosomal maturation, autophagy, production of cytokines, chemokines and anti-microbial peptides, and functional characterization of individual cell types. Strikingly, the in vitro tissue model allows manipulation of one or more cells types and provides a relevant tissue micro-environment, not only for studies on TB, but for a variety of infectious and non-infectious diseases that affect the lungs.

ヒトでは、感染症、組織炎症、細胞の動員、組織リモデリングおよび組織の恒常性の調節に応答が異なる細胞型が関与する複雑な事象です。したがって、これらの方法は、最高の局所組織環境において研究されています。以前は、これは、主に実験動物モデルを使用して可能でした。彼らはしばしば、人間とは異なる方法で病原体に対応し、また、疾患1の別のコースを表示しかし、広く使用されている実験動物は、多くの制限を保持します。 インビトロの肺組織モデル中のヒトはヒト肺で特異的免疫応答を研究するための可能性を保持しています。 ヒト結核感染（TB）は、主に肺に影響を与える病気である。 結核菌 （ 結核菌 ）、結核の原因物質は、細菌が肺dendriに飲み込まれている肺胞腔に輸送されるエアロゾル液滴を経由して肺に達します感染2,3に対する自然免疫応答の一部として、チック細胞や肺胞マクロファージ。病原体の食作用はファゴソーム内のバグの区画化につながる、理想的には食細胞による中和および病原体を死滅させることになります。 Mに曝露された個体の50％まで結核は、先天性免疫応答4を介して感染をクリアすることができると考えられています。感染の他の結果は後の段階で適応免疫系によるクリアランス、潜伏感染またはアクティブな疾患5慢性最悪のケースです。 以前は人間の結核の研究のために何のインビトロ組織モデルは存在していません。ヒトマクロファージまたは他の末梢血細胞の単一細胞培養物は、多くの場合、6,7を使用されています。このアプローチの欠点は、Mに露出し、肺組織内で一緒に動作して、異なる細胞型の動態を反映することができないことです結核 </eM&gt;。したがって、TB上の機能とメカニズムの研究を行うことができるようにするインビトロモデルが必要とされています。セル・ベースのここで説明する体外ヒト肺組織モデルでは、もともとの樹状細胞の機能8の研究のために私たちのグループによって設立されました。私たちは、結核の研究のために、この方法を適応しています。 ここに提示されたヒト肺組織モデルは、組織特異的な上皮細胞および線維芽細胞8から成ります。これらの細胞は、トランスウェルインサート中の多孔質膜の上に、コラーゲンマトリックス中で培養され、フォーム構造は、正常なヒトの肺組織（ 図1）に似ています。空気にさらされると細胞は頂端側8に粘液を分泌し始めます。 M.に感染したヒト初代マクロファージを注入することにより、モデルへの結核 、我々は、免疫細胞が組織に移行し、結核性肉芽腫9の初期段階を形成する方法を観察しています。これは最初のヒト組織モデルDESCRです結核ibed、それが結核と肺の他の疾患に対する自然免疫応答を研究するための有望なツールをもたらします。これまでに、我々は、モデルにおいて免疫細胞としてのみ単球およびマクロファージを使用しているが、複雑さのレベルは、追加の関連する細胞型を含めることによって増加させることができます。 <img alt="図1" src="/files/ftp_upload/53084/53084fig1.jpg" /> 肺組織モデルの概略1.概略図。 （A）モデルM.、ヒト肺特異的上皮細胞で構成されています結核は、初代マクロファージをに感染し、赤色の色素ラベルされた単球をトランスウェルフィルター上に作製したコラーゲン埋め込 ​​まれた線維芽細胞上に播種。空気への組織モデルの露出は、上皮によって細胞外マトリックスタンパク質の産生、粘液分泌と層化を開始します。こうして開発された3次元組織モデルは 、M を研究するための便利なツールですCLO環境における結核感染SELYヒトの肺。（B）組織モデルの製造における様々なステップの代表的な顕微鏡像。（C）肺モデル組織切片の完全な構造に似ています。スケール- 。100ミクロン<a href="https://www.jove.com/files/ftp_upload/53084/53084fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a>

<table><tbody><tr><td>Cell culture inserts</td><td>BD Falcon</td><td>353092</td><td></td></tr><tr><td>6-well culture plates</td><td>BD Falcon</td><td>353046</td><td></td></tr><tr><td>MRC-5 cells, lung fibroblasts</td><td></td><td></td><td>ATCC#CCL-171</td></tr><tr><td>16HBE cells, lung epithelial cells</td><td></td><td></td><td>Gift from Dr. Dieter Gruenert, Mt. Zion Cancer Center, University of California, San Fransisco, USA&amp;nbsp;</td></tr><tr><td>5 x Dulbecco&amp;rsquo;s modified Eagle&amp;rsquo;s medium (5 x DMEM)</td><td>Gibco</td><td>12800-082</td><td>Made from powder but add 5 times less water. Adjust pH to 7.3 and filter it using a 0.2 &amp;micro;m filter.</td></tr><tr><td>Dulbecco&amp;rsquo;s modified Eagle&amp;rsquo;s medium with glucose (DMEM) 1x</td><td>Gibco</td><td>41965-039</td><td></td></tr><tr><td>Minimum Essential Medium (MEM) 1x with Earle&amp;rsquo;s salts</td><td>Sigma</td><td>M4655</td><td></td></tr><tr><td>Non-Essential Amino Acids Solution, 100x</td><td>Life Technologies</td><td>11140-035</td><td></td></tr><tr><td>L-glutamine 200 mM (100x)</td><td>Gibco</td><td>25030-024</td><td></td></tr><tr><td>Sodium Pyruvate</td><td>Life Technologies</td><td>11360-039</td><td></td></tr><tr><td>NaHCO3 (71.2 mg/ml)</td><td></td><td></td><td>Prepared in house</td></tr><tr><td>Heat inactivated Fetal Bovine Serum (FBS)</td><td>Gibco</td><td>10270-106</td><td>Heat inactivated for 30 min, 56 &amp;deg;C</td></tr><tr><td>Gentamicin (50 mg/ml)</td><td>Gibco</td><td>15750-060</td><td></td></tr><tr><td>Hepes buffer solution 1M</td><td>Gibco</td><td>15630-056</td><td></td></tr><tr><td>Penicillin Streptomycin (Pen Strep)</td><td>Gibco</td><td>15140-122</td><td></td></tr><tr><td>Lymphoprep</td><td>Axis-Shield</td><td>7801</td><td></td></tr><tr><td>Ultrapure 0.5 M EDTA</td><td>Gibco</td><td>15575</td><td></td></tr><tr><td>Bovine Collagen PA treated (500 ml)</td><td>Organogenesis</td><td>200-055</td><td></td></tr><tr><td>Pure col purified Bovine Collagen solution (100 ml)</td><td>Advanced biomatrix</td><td>5005-B</td><td></td></tr><tr><td>Extracellular matrix protein, Fibronectin (1 mg)</td><td>BD</td><td>354008</td><td></td></tr><tr><td>Primary human monocytes/macrophages</td><td></td><td></td><td>Isolated from human whole blood or buffy coats.</td></tr><tr><td>PKH26 Red fluorescent cell linker</td><td>Sigma</td><td>MINI26</td><td></td></tr><tr><td>Mycobacterium tuberculosis H37Rv expressing green fluorescent protein</td><td></td><td></td><td>M. tuberculosis H37Rv wild type was transformed with the pFPV2 plasmid constitutively expressing GFP.</td></tr><tr><td>Middlebrook 7H9 medium&amp;nbsp;</td><td>Difco</td><td>271310</td><td></td></tr><tr><td>BBL Middlebrook ADC Enrichment</td><td>BBL</td><td>211887</td><td></td></tr><tr><td>Tween-80</td><td></td><td></td><td></td></tr><tr><td>Glycerol</td><td></td><td></td><td></td></tr><tr><td>Kanamycin B sulfate (20 &amp;micro;g/ml)</td><td>Sigma</td><td>B5264</td><td></td></tr><tr><td>Prolong Gold anti=-fade reagent with DAPI</td><td>Invitrogen</td><td>P36935</td><td></td></tr><tr><td>Trypsin -EDTA</td><td></td><td></td><td></td></tr><tr><td>Bovine serum albumin</td><td></td><td></td><td></td></tr><tr><td>Paraformaldehyde</td><td></td><td></td><td></td></tr><tr><td>DAPI</td><td></td><td></td><td></td></tr><tr><td>LSM700 Confocal microscope</td><td>Zeiss</td><td></td><td></td></tr><tr><td>iMaris Scientific 3D/4D image processing software, version 7.6.8</td><td>Bitplane AG</td><td></td><td></td></tr></tbody></table>

a 3d human lung tissue model for functional studies on mycobacterium tuberculosis infection

結核（TB）は依然として世界の人々の健康に大きな脅威を保持し、私たちは病気のメカニズムを理解し、新たな治療選択肢の発見を進める手助けするためのコスト効率に優れたが、信頼性の高いモデルの必要性がある。単層のインビトロ細胞培養または共培養は、三次元（3D）環境および組織応答を欠いています。ここで、我々は結核菌 （ 結核菌 ）の感染の間に起こる複雑な事象を研究するための有効なツールであることが期待されているヒト肺組織のインビトロモデルで革新的なについて説明します。 3次元組織モデルは、多孔質膜の上に、コラーゲンマトリックス中で培養された組織特異的上皮細胞および線維芽細胞からなります。空気暴露されると、上皮細胞層別化し、頂端側の粘液を分泌します。 M.に感染したヒト初代マクロファージを導入することにより組織モードに結核lは、我々は、免疫細胞が感染組織中に移動し、TBの肉芽腫の初期段階を形成することが示されています。これらの構造は根本的に異なるか、一般的に広く使用されている実験動物モデルにおいて観察されていない人間の結核、肉芽腫の明確な特徴を再現。この器官培養法は、宿主細胞病原体相互作用の空間的および時間的機能に関する重要な情報を提供する3次元可視化と堅牢な定量分析を可能にします。まとめると、肺組織モデルは、結核の研究のための生理学的に関連する組織の微小環境を提供します。従って、肺組織モデルは、基本的な機構的および応用研究の両方のための潜在的な意味を持ちます。重要なことには、モデルは、それによって、肺に影響を与える感染症の様々なモデル化するためのその使用が広がり、個々の細胞型の添加または操作を可能にします。

ヒトの結核のための3D肺組織モデルは、効果的にMの宿主-病原体相互作用を研究するために使用することができ結核感染 。この方法の基本的な手順は、異なるステップ及び組織切片の全体的な微細構造の代表的な顕微鏡写真を図1に概説されている。このモデルは肺線維芽細胞、気管支上皮細胞および初代単球/マクロファージを含むヒト肺組織のいくつかの構成要素を有しています3D組織環境内に埋め込まれました。ヒト肺組織の構成要素を組み込むだけでなく、モデルは、生理的条件に上皮および粘液分泌のすなわち成層に似ています。 TB感染の監視に肺組織モデルの使用の例は、図2に示されている。M.を視覚化するため結核 -immune細胞遊走との相互作用は、我々 は 、Mに感染したマクロファージを導入しましたGFPを発現する結核組織モデル（核について染色ブルー、DAPI）に合わせて新たに単離したPKH26標識単球（赤）と（緑）。 Mの 7日後の追加について組織モデルに結核に感染した細胞は、共焦点顕微鏡は、人間の結核9の特徴的な病変を模倣感染（緑）のサイト（ 図2）、で赤単球のクラスターを明らかにしています。 Mの 3次元可視化のための代表的な一連の画像結核は、細胞クラスターの組織モデルと定量化は、図3に示されているに感染し。3Dビジュアライゼーションは、3D画像にいくつかの機能を、対話調べ、定量化するため、ユーザーの柔軟性を提供します。 M.部位で単球のクラスターを明らかに図3Bに示すように、緑と赤の細菌単球クラスタの空間的配置は、頂端の回転及び側面図から分かるよう結核。クラスター OBませんでした感染していない組織 （ 図3A）で提供しています。個々の単球の数がMに（p &lt;0.01）減少し、我々は、単球細胞集団のサイズと数を定量した （p &lt;0.001）、細胞塊の大きさ（体積）が増強されることを見出しました結核は感染していない組織モデル（ 図3Cおよび3D）と比較して組織を感染させました。このデータは、Mの初期の肉芽腫形成の我々の以前の発見を検証します結核感染は、2D組織切片9で分析肺組織モデルで観察しました。 我々のデータは、組織モデルは、M細胞-複雑なホストを調査するために自然な立体生息地を提供していることを示唆しています結核の通信ネットワーク。また、3次元可視化と定量分析は、組織モデル（ 図3）の機能を研究するための優れたツールであることがわかりました。細胞塊の定量化（例えば、肉芽腫）、多くの場合、ストレッチ<html>いくつかの細胞層のHESとは完全に3次元定量分析によって捕捉することができます。また、モデル内の個々の細胞や細菌の正確な空間的および時間的特徴の可視化は、指定された実験室でのライブイメージング、移動および追跡調査を可能にします。 <img alt="図2" src="/files/ftp_upload/53084/53084fig2.jpg" /> 病原性M.周囲の組織モデルクラスター図2.単球結核。感染していないと、Mの代表的な共焦点画像結核に感染した組織モデルが提示されています。感染していない組織と比較して、パネルの緑色から（M. tuberculosis- GFP）、赤（PKH26標識単球）、青（DAPI染色核）と合併チャネルが感染組織における単球の動員を示します。スケール - 100μmです。large.jpg &quot;スタイル=&quot;フォントサイズ：14px;行の高さ：28px; &quot;ターゲット=&quot; _空白 &quot;&gt;この図の拡大版をご覧になるにはこちらをクリックしてください。 <img alt="図3" src="/files/ftp_upload/53084/53084fig3.jpg" /> 図3. 3次元可視化と組織モデルの定量分析は、有用な情報を提供しています。全体の組織モデル（A）非感染組織の3次元可視化の代表的な画像、（B）に感染したM.結核、IMARIS画像処理ソフトウェア（バージョン7.6.8）によりツァイスLSM700共焦点顕微鏡および定量分析を用いた光学セクショニングを介し。これらの画像は20X倍率で取得された、頂端、回転水平、回転、垂直および側面図（AとB）からの可視化を可能にする、1.5μmの間隔で19.5ミクロンの組織の厚さをカバーする14のzスタック。（C）QUA単球細胞クラスターのntitative分析はM.後早期肉芽腫クラスタの増強した（p &lt;0.0001）のサイズを明らかに感染組織で複数のクラスタを繰り返し強調、感染していない組織と比較して、感染の不在に比べ結核感染 。（D）単球数の定量は、感染組織の減少した（p &lt;0.01）を示しました。グリーン- M.結核 -GFP、レッド- PKH26標識単球、ブルー-細胞核、スケール- 。100ミクロン<a href="https://www.jove.com/files/ftp_upload/53084/53084fig3large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a>

Watch this Scientific Journal Video about A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection at JoVE.com

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

Human tuberculosis infection is a complex process, which is difficult to model in vitro. Here we describe a novel 3D human lung tissue model that recapitulates the dynamics that occur during infection, including the migration of immune cells and early granuloma formation in a physiological environment.

上の機能的研究のための3Dヒト肺組織モデル<em&gt;結核菌</em&gt;感染

Tuberculosis (TB) still holds a major threat to the health of people worldwide, and there is a need for cost-efficient but reliable models to help us ...

a-3d-human-lung-tissue-model-for-functional-studies-on-mycobacterium

Research

JoVE Journal

Immunology and Infection

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

18.9K ビュー.  Linköping University. Human tuberculosis infection is a complex process, which is difficult to model in vitro. Here we describe a novel 3D human lung tissue model that recapitulates the dynamics that occur during infection, including the migration of immune cells and early granuloma formation in a physiological environment.

ビデオ: A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

An Experimental Model to Study Tuberculosis-Malaria Coinfection upon Natural Transmission of Mycobacterium tuberculosis and Plasmodium berghei

Coinfections naturally occur due to the geographic overlap of distinct types of pathogenic organisms. Concurrent infections most likely modulate the respective immune response to each single pathogen and may thereby affect pathogenesis and disease outcome. Coinfected patients may also respond differentially to anti-infective interventions. Coinfection between tuberculosis as caused by mycobacteria and the malaria parasite Plasmodium, both of which are coendemic in many parts of sub-Saharan Africa, has not been studied in detail. In order to approach the challenging but scientifically and clinically highly relevant question how malaria-tuberculosis coinfection modulate host immunity and the course of each disease, we established an experimental mouse model that allows us to dissect the elicited immune responses to both pathogens in the coinfected host. Of note, in order to most precisely mimic naturally acquired human infections, we perform experimental infections of mice with both pathogens by their natural routes of infection, i.e. aerosol and mosquito bite, respectively.

An Experimental Model to Study Tuberculosis-Malaria Coinfection upon Natural Transmission of Mycobacterium tuberculosis and Plasmodium berghei

Tuberculosis and malaria are two of the most prevalent infections in humans and major causes of morbidity and mortality in impoverished populations in the tropics. We established an experimental model system to study outcome of malaria-tuberculosis coinfection in mice after challenge with both pathogens via their natural route of infection.

自然送信時に結核マラリア重複感染を研究する実験モデル結核菌とマラリアベルゲイ</i

Coinfections naturally occur due to the geographic overlap of distinct types of pathogenic organisms. Concurrent infections most likely modulate the ...

免疫・感染学

A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds

In the present study, we combined an in vitro 3D lung tumor model with an in silico model to optimize predictions of drug response based on a specific mutational background. The model is generated on a decellularized porcine scaffold that reproduces tissue-specific characteristics regarding extracellular matrix composition and architecture including the basement membrane. We standardized a protocol that allows artificial tumor tissue generation within 14 days including three days of drug treatment. Our article provides several detailed descriptions of 3D read-out screening techniques like the determination of the proliferation index Ki67 staining&#39;s, apoptosis from supernatants by M30-ELISA and assessment of epithelial to mesenchymal transition (EMT), which are helpful tools for evaluating the effectiveness of therapeutic compounds. We could show compared to 2D culture a reduction of proliferation in our 3D tumor model that is related to the clinical situation. Despite of this lower proliferation, the model predicted EGFR-targeted drug responses correctly according to the biomarker status as shown by comparison of the lung carcinoma cell lines HCC827 (EGFR -mutated, KRAS wild-type) and A549 (EGFR wild-type, KRAS-mutated) treated with the tyrosine-kinase inhibitor (TKI) gefitinib. To investigate drug responses of more advanced tumor cells, we induced EMT by long-term treatment with TGF-beta-1 as assessed by vimentin/pan-cytokeratin immunofluorescence staining. A flow-bioreactor was employed to adjust culture to physiological conditions, which improved tissue generation. Furthermore, we show the integration of drug responses upon gefitinib treatment or TGF-beta-1 stimulation - apoptosis, proliferation index and EMT - into a Boolean in silico model. Additionally, we explain how drug responses of tumor cells with a specific mutational background and counterstrategies against resistance can be predicted. We are confident that our 3D in vitro approach especially with its in silico expansion provides an additional value for preclinical drug testing in more realistic conditions than in 2D cell culture.

A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds

We present a three-dimensional (3D) lung cancer model based on a biological collagen scaffold to study sensitivity towards non-small-cell-lung-cancer-(NSCLC)-targeted therapies. We demonstrate different read-out techniques to determine the proliferation index, apoptosis and epithelial-mesenchymal transition (EMT) status. Collected data are integrated into an in silico model for prediction of drug sensitivity.

操作された複合三次元組織<em&gt;インビトロ</em&gt; /<em&gt;インシリコ</em具体的な突然変異背景に薬物の有効性を予測するための&gt;肺腫瘍モデル

In the present study, we combined an in vitro 3D lung tumor model with an in silico model to optimize predictions of drug response based on a specific ...

生物工学

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a leading cause of morbidity and mortality worldwide. With the increased spread of multi drug-resistant TB (MDR-TB), there is a real urgency to develop new therapeutic strategies against M. tuberculosis infections. Traditionally, compounds are evaluated based on their antibacterial activity under in vitro growth conditions in broth; however, results are often misleading for intracellular pathogens like M. tuberculosis since in-broth phenotypic screening conditions are significantly different from the actual disease conditions within the human body. Screening for inhibitors that work inside macrophages has been traditionally difficult due to the complexity, variability in infection, and slow replication rate of M. tuberculosis. In this study, we report a new approach to rapidly assess the effectiveness of compounds on the viability of M. tuberculosis in a macrophage infection model. Using a combination of a cytotoxicity assay and an in-broth M. tuberculosis viability assay, we were able to create a screening system that generates a comprehensive analysis of compounds of interest. This system is capable of producing quantitative data at a low cost that is within reach of most labs and yet is highly scalable to fit large industrial settings.

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

We have developed a modular high-throughput screening system for discovering novel compounds against Mycobacterium tuberculosis, targeting intracellular and in-broth growing bacteria as well as cytotoxicity against the mammalian host cell.

細胞内の標的阻害剤の有効性及び細胞毒性スクリーニングのためのシステム<em&gt;結核菌</em

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a leading cause of morbidity and mortality worldwide. With the increased spread ...

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

Reporter enzyme fluorescence (REF) utilizes substrates that are specific for enzymes present in target organisms of interest for imaging or detection by fluorescence or bioluminescence. We utilize BlaC, an enzyme expressed constitutively by all M. tuberculosis strains. REF allows rapid quantification of bacteria in lungs of infected mice. The same group of mice can be imaged at many time points, greatly reducing costs, enumerating bacteria more quickly, allowing novel observations in host-pathogen interactions, and increasing statistical power, since more animals per group are readily maintained. REF is extremely sensitive due to the catalytic nature of the BlaC enzymatic reporter and specific due to the custom flourescence resonance energy transfer (FRET) or fluorogenic substrates used. REF does not require recombinant strains, ensuring normal host-pathogen interactions. We describe the imaging of M. tuberculosis infection using a FRET substrate with maximal emission at 800 nm. The wavelength of the substrate allows sensitive deep tissue imaging in mammals. We will outline aerosol infection of mice with M. tuberculosis, anesthesia of mice, administration of the REF substrate, and optical imaging. This method has been successfully applied to evaluating host-pathogen interactions and efficacy of antibiotics targeting M. tuberculosis.

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

We describe the optical imaging of mice infected with Mycobacterium tuberculosis (M. tuberculosis) using reporter enzyme fluorescence (REF). This protocol facilitates the sensitive and specific detection of M. tuberculosis in pre-clinical animal models for pathogenesis, therapeutics and vaccine research.

レポーター酵素蛍光を用いたマウスにおける結核菌をイメージング

Reporter enzyme fluorescence (REF) utilizes substrates that are specific for enzymes present in target organisms of interest for imaging or detection by ...

Modeling Tuberculosis in Mycobacterium marinum Infected Adult Zebrafish

Mycobacterium tuberculosis is currently the deadliest human pathogen causing 1.7 million deaths and 10.4 million infections every year. Exposure to this bacterium causes a wide disease spectrum in humans ranging from a sterilized infection to an actively progressing deadly disease. The most common form is the latent tuberculosis, which is asymptomatic, but has the potential to reactivate into a fulminant disease. Adult zebrafish and its natural pathogen Mycobacterium marinum have recently proven to be an applicable model to study the wide disease spectrum of tuberculosis. Importantly, spontaneous latency and reactivation as well as adaptive immune responses in the context of mycobacterial infection can be studied in this model. In this article, we describe methods for the experimental infection of adult zebrafish, the collection of internal organs for the extraction of nucleic acids for the measurement of mycobacterial loads and host immune responses by quantitative PCR. The in-house-developed, M. marinum-specific qPCR assay is more sensitive than the traditional plating methods as it also detects DNA from non-dividing, dormant or recently dead mycobacteria. As both DNA and RNA are extracted from the same individual, it is possible to study the relationships between the diseased state, and the host and pathogen gene-expression. The adult zebrafish model for tuberculosis thus presents itself as a highly applicable, non-mammalian in vivo system to study host-pathogen interactions.

Modeling Tuberculosis in Mycobacterium marinum Infected Adult Zebrafish

Here, we present a protocol to model human tuberculosis in an adult zebrafish using its natural pathogen Mycobacterium marinum. Extracted DNA and RNA from the internal organs of infected zebrafish can be used to reveal the total mycobacterial loads in the fish and the host&#39;s immune responses with qPCR.

結核抗酸菌 marinum感染成人ゼブラフィッシュをモデル化

Mycobacterium tuberculosis is currently the deadliest human pathogen causing 1.7 million deaths and 10.4 million infections every year. Exposure to this ...

Evaluating Regional Pulmonary Deposition using Patient-Specific 3D Printed Lung Models

Development of targeted therapies for pulmonary diseases is limited by the availability of preclinical testing methods with the ability to predict regional aerosol delivery. Leveraging 3D printing to generate patient-specific lung models, we outline the design of a high-throughput, in vitro experimental setup for quantifying lobular pulmonary deposition. This system is made with a combination of commercially available and 3D printed components and allows the flow rate through each lobe of the lung to be independently controlled. Delivery of fluorescent aerosols to each lobe is measured using fluorescence microscopy. This protocol has the potential to promote the growth of personalized medicine for respiratory diseases through its ability to model a wide range of patient demographics and disease states. Both the geometry of the 3D printed lung model and the air flow profile setting can be easily modulated to reflect clinical data for patients with varying age, race, and gender. Clinically relevant drug delivery devices, such as the endotracheal tube shown here, can be incorporated into the testing setup to more accurately predict a device&#8217;s capacity to target therapeutic delivery to a diseased region of the lung. The versatility of this experimental setup allows it to be customized to reflect a multitude of inhalation conditions, enhancing the rigor of preclinical therapeutic testing.

We present a high-throughput, in vitro method for quantifying regional pulmonary deposition at the lobe level using CT scan-derived, 3D printed lung models with tunable air flow profiles.

患者固有の3Dプリント肺モデルを用いた局所肺堆積の評価

Development of targeted therapies for pulmonary diseases is limited by the availability of preclinical testing methods with the ability to predict ...

Use of the Invertebrate Galleria mellonella as an Infection Model to Study the Mycobacterium tuberculosis Complex

Tuberculosis is the leading global cause of infectious disease mortality and roughly a quarter of the world&#8217;s population is believed to be infected with Mycobacterium tuberculosis. Despite decades of research, many of the mechanisms behind the success of M. tuberculosis as a pathogenic organism remain to be investigated, and the development of safer, more effective antimycobacterial drugs are urgently needed to tackle the rise and spread of drug resistant tuberculosis. However, the progression of tuberculosis research is bottlenecked by traditional mammalian infection models that are expensive, time consuming, and ethically challenging. Previously we established the larvae of the insect Galleria mellonella (greater wax moth) as a novel, reproducible, low cost, high-throughput and ethically acceptable infection model for members of the M. tuberculosis complex. Here we describe the maintenance, preparation, and infection of G. mellonella with bioluminescent Mycobacterium bovis BCG lux. Using this infection model, mycobacterial dose dependent virulence can be observed, and a rapid readout of in vivo mycobacterial burden using bioluminescence measurements is easily achievable and reproducible. Although limitations exist, such as the lack of a fully annotated genome for transcriptomic analysis, ontological analysis against genetically similar insects can be carried out. As a low cost, rapid, and ethically acceptable model for tuberculosis, G. mellonella can be used as a pre-screen to determine drug efficacy and toxicity, and to determine comparative mycobacterial virulence prior to the use of conventional mammalian models. The use of the G. mellonella-mycobacteria model will lead to a reduction in the substantial number of animals currently used in tuberculosis research.

Use of the Invertebrate Galleria mellonella as an Infection Model to Study the Mycobacterium tuberculosis Complex

Galleria mellonella was recently established as a reproducible, cheap, and ethically acceptable infection model for the Mycobacterium tuberculosis complex. Here we describe and demonstrate the steps taken to establish successful infection of G. mellonella with bioluminescent Mycobacterium bovis BCG lux.

結核複合体を研究するための感染モデルとしての無脊椎動物ガレリアメロネラの使用

Tuberculosis is the leading global cause of infectious disease mortality and roughly a quarter of the world&#8217;s population is believed to be infected ...

Spatial Quantification of Drugs in Pulmonary Tuberculosis Lesions by Laser Capture Microdissection Liquid Chromatography Mass Spectrometry (LCM-LC/MS)

Tuberculosis is still a leading cause of morbidity and mortality worldwide. Improvements to existing drug regimens and the development of novel therapeutics are urgently required. The ability of dosed TB drugs to reach and sterilize bacteria within poorly-vascularized necrotic regions (caseum) of pulmonary granulomas is crucial for successful therapeutic intervention. Effective therapeutic regimens must therefore contain drugs with favorable caseum penetration properties. Current LC/MS methods for quantifying drug levels in biological tissues have limited spatial resolution capabilities, making it difficult to accurately determine absolute drug concentrations within small tissue compartments such as those found within necrotic granulomas. Here we present a protocol combining laser capture microdissection (LCM) of pathologically-distinct tissue regions with LC/MS quantification. This technique provides absolute quantification of drugs within granuloma caseum, surrounding cellular lesion and uninvolved lung tissue and, therefore, accurately determines whether bactericidal concentrations are being achieved. In addition to tuberculosis research, the technique has many potential applications for spatially-resolved quantification of drugs in diseased tissues.

Spatial Quantification of Drugs in Pulmonary Tuberculosis Lesions by Laser Capture Microdissection Liquid Chromatography Mass Spectrometry LCM-LC/MS

Here, we describe a protocol using laser capture microdissection coupled with LC/MS analysis to spatially-quantify drug distributions within pulmonary tuberculosis granulomas. The approach has broad applicability to quantifying drug concentrations within tissues at high spatial detail.

レーザーによる肺の結核病変における薬物の空間の定量化をキャプチャ レーザーマイクロダイ セクション液体クロマトグラフィー質量分析法 (LCM-LC/MS)

Tuberculosis is still a leading cause of morbidity and mortality worldwide. Improvements to existing drug regimens and the development of novel ...

医学

P. aeruginosa Infected 3D Co-Culture of Bronchial Epithelial Cells and Macrophages at Air-Liquid Interface for Preclinical Evaluation of Anti-Infectives

fDrug research for the treatment of lung infections is progressing towards predictive in vitro models of high complexity. The multifaceted presence of bacteria in lung models can re-adapt epithelial arrangement, while immune cells coordinate an inflammatory response against the bacteria in the microenvironment. While in vivo models have been the choice for testing new anti-infectives in the context of cystic fibrosis, they still do not accurately mimic the in vivo conditions of such diseases in humans and the treatment outcomes. Complex in vitro models of the infected airways based on human cells (bronchial epithelial and macrophages) and relevant pathogens could bridge this gap and facilitate the translation of new anti-infectives into the clinic. For such purposes, a co-culture model of the human cystic fibrosis bronchial epithelial cell line CFBE41o- and THP-1 monocyte-derived macrophages has been established, mimicking an infection of the human bronchial mucosa by P. aeruginosa at air-liquid interface (ALI) conditions. This model is set up in seven days, and the following parameters are simultaneously assessed: epithelial barrier integrity, macrophage transmigration, bacterial survival, and inflammation. The present protocol describes a robust and reproducible system for evaluating drug efficacy and host responses that could be relevant for discovering new anti-infectives and optimizing their aerosol delivery to the lungs.

P. aeruginosa Infected 3D Co-Culture of Bronchial Epithelial Cells and Macrophages at Air-Liquid Interface for Preclinical Evaluation of Anti-Infectives

We describe a protocol for a three-dimensional co-culture model of infected airways, using CFBE41o- cells, THP-1 macrophages, and Pseudomonas aeruginosa, established at the air-liquid interface. This model provides a new platform to simultaneously test antibiotic efficacy, epithelial barrier function, and inflammatory markers.

P. アフルギノーサ 抗感染性の前臨床評価のための空気・液体界面における気管支上皮細胞とマクロファージの感染3Dコカルチャー

fDrug research for the treatment of lung infections is progressing towards predictive in vitro models of high complexity. The multifaceted presence of ...

Generation of Human 3D Lung Tissue Cultures (3D-LTCs) for Disease Modeling

Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) used as 3D lung tissue cultures (3D-LTCs) represent an elegant and biologically highly relevant 3D cell culture model, which highly resemble in situ tissue due to their complexity, biomechanics and molecular composition. Tissue slicing is widely applied in various animal models. 3D-LTCs derived from human PCLS can be used to analyze responses to novel drugs, which might further help to better understand the mechanisms and functional effects of drugs in human tissue. The preparation of PCLS from surgically resected lung tissue samples of patients, who experienced lung lobectomy, increases the accessibility of diseased and peritumoral tissue. Here, we describe a detailed protocol for the generation of human PCLS from surgically resected soft-elastic patient lung tissue. Agarose was introduced into the bronchoalveolar space of the resectates, thus preserving lung structure and increasing the tissue&#39;s stiffness, which is crucial for subsequent slicing. 500 &#181;m thick slices were prepared from the tissue block with a vibratome. Biopsy punches taken from PCLS ensure comparable tissue sample sizes and further increase the amount of tissue samples. The generated lung tissue cultures can be applied in a variety of studies in human lung biology, including the pathophysiology and mechanisms of different diseases, such as fibrotic processes at its best at (sub-)cellular levels. The highest benefit of the 3D-LTC ex vivo model is its close representation of the in situ human lung in respect of 3D tissue architecture, cell type diversity and lung anatomy as well as the potential for assessment of tissue from individual patients, which is relevant to further develop novel strategies for precision medicine.

Generation of Human 3D Lung Tissue Cultures 3D-LTCs for Disease Modeling

Here, we present a protocol for the preparation of agarose-filled human precision-cut lung slices from resected patient tissue that are suitable for generating 3D lung tissue cultures to model human lung diseases in biological and biomedical studies.

疾患モデル作製のひと 3 D 肺組織培養 (3 D-Ltc) の生成

Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) ...

上の機能的研究のための3Dヒト肺組織モデル

要約

さらに動画を探す

この動画の章

自然送信時に結核マラリア重複感染を研究する実験モデル結核菌とマラリアベルゲイ

操作された複合三次元組織

細胞内の標的阻害剤の有効性及び細胞毒性スクリーニングのためのシステム

レポーター酵素蛍光を用いたマウスにおける結核菌をイメージング

結核抗酸菌 marinum感染成人ゼブラフィッシュをモデル化

患者固有の3Dプリント肺モデルを用いた局所肺堆積の評価

結核複合体を研究するための感染モデルとしての無脊椎動物ガレリアメロネラの使用

レーザーによる肺の結核病変における薬物の空間の定量化をキャプチャレーザーマイクロダイセクション液体クロマトグラフィー質量分析法 (LCM-LC/MS)

P. アフルギノーサ抗感染性の前臨床評価のための空気・液体界面における気管支上皮細胞とマクロファージの感染3Dコカルチャー

疾患モデル作製のひと 3 D 肺組織培養 (3 D-Ltc) の生成

上の機能的研究のための3Dヒト肺組織モデル

要約

さらに動画を探す

この動画の章

自然送信時に結核マラリア重複感染を研究する実験モデル結核菌とマラリアベルゲイ

操作された複合三次元組織

細胞内の標的阻害剤の有効性及び細胞毒性スクリーニングのためのシステム

レポーター酵素蛍光を用いたマウスにおける結核菌をイメージング

結核抗酸菌 marinum感染成人ゼブラフィッシュをモデル化

患者固有の3Dプリント肺モデルを用いた局所肺堆積の評価

結核複合体を研究するための感染モデルとしての無脊椎動物ガレリアメロネラの使用

レーザーによる肺の結核病変における薬物の空間の定量化をキャプチャ レーザーマイクロダイ セクション液体クロマトグラフィー質量分析法 (LCM-LC/MS)

P. アフルギノーサ 抗感染性の前臨床評価のための空気・液体界面における気管支上皮細胞とマクロファージの感染3Dコカルチャー

疾患モデル作製のひと 3 D 肺組織培養 (3 D-Ltc) の生成

レーザーによる肺の結核病変における薬物の空間の定量化をキャプチャレーザーマイクロダイセクション液体クロマトグラフィー質量分析法 (LCM-LC/MS)

P. アフルギノーサ抗感染性の前臨床評価のための空気・液体界面における気管支上皮細胞とマクロファージの感染3Dコカルチャー