Name: a standardized method for measuring internal lung surface area via mouse pneumonectomy and prosthesis implantation
Uploaded: 2017-07-26T15:00:00
Description: Watch this Scientific Journal Video about A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation at JoVE.com

Forfatterne vil gerne anerkende National Institute of Biological Sciences, Beijing for at få hjælp. Dette arbejde blev støttet af Beijing Municipal Natural Science Foundation (nr. Z17110200040000).

Alle procedurer anvendt i denne protokol blev udført i overensstemmelse med anbefalingerne i retningslinjerne for pleje og brug af laboratoriedyr fra National Institute of Biological Sciences, Beijing. 8 ugers gamle CD-1 hanmus blev anbragt i en specifik patogenfri (SPF) facilitet, indtil forsøgene blev udført. Surgerier blev udført ved hjælp af fuldstændigt bedøvede mus ( dvs. uden tåbspidsreaktioner). Efter operationen blev musene holdt i et varmt, fugtigt rum med tilstrækkelig mad og ferskvand. Mus ...

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I denne protokol giver vi detaljerede beskrivelser af måling af pulmonale parametre efter musen venstre lunge PNX og protese implantation. ISA anses nu for at være en vigtig metrisk til vurdering af åndedrætsfunktionen i mange lungesygdomme og ved skadefremkaldt alveolær regenerering. Selv om det lungeforskningsmiljø er enig om brugen af ​​ISA som en nyttig metrisk, har der indtil videre været ringe overvejelse om standardiseringen af ​​måling af ILV og MLI, de to parametre, der bruges til at beregne ISA...

Lungens grundlæggende funktion er udvekslingen af ​​ilt og kuldioxid mellem blodkar og atmosfære. Lungesygdomme såsom bronkopulmonær dysplasi (BPD), kronisk obstruktiv lungesygdom (COPD) og akut respiratoriske infektioner resulterer i nedsat ISA 2 . Forskere, der studerer lungesygdomme, har udviklet flere kvantitative metoder til evaluering af morfologiske ændringer i lungerne, herunder MLI, ILV, antal gasudvekslingsenheder, ISA og lungevævsoverensstemmelse 2 , 3 . Banebrydende undersøgelser af Weibel et al. 4 og Duguid et al. 5 fastslog sammen, at ISA kan anvendes som en direkte måling af lungegasudvekslingskapacitet i menneskelige lunger og kan anvendes som et kriterium for at bestemme sværhedsgraden af ​​emfysem. En række undersøgelser udgivet i de sidste fem år har brugt lungemorfologiske parametre ( f.eks. </eM&gt; ISA og MLI) for at vurdere morfologiske og funktionelle ændringer i muskelungen under udvikling 6 og under genopretning fra skade PNX 1 , 7 . ISA beregnes ved anvendelse af ligning 1 8 , 9 : <img alt="ligning" src="/files/ftp_upload/56114/56114eq1.jpg" /> , Hvor ILV er det interne lungemængde, og MLI er en mellemliggende parameter, der repræsenterer den lunge perifere luftrumsstørrelse 10 . PNX, den kirurgiske fjernelse af en eller flere lungelaber, er blevet rapporteret meget for at inducere alveolær regenerering hos mange arter, herunder mennesker 11 , mus 1 , hunde 12 , rotter 13 og kaniner 14 , 15 . En studY af mus lunger på fjorten dage efter PNX viste, at både ekspansion af eksisterende alveoler og de novo dannelsen af ​​alveoler bidrager til genoprettelsen af ​​ISA, ILV og antallet af alveoler i de resterende lungevæv 1 . Vi og andre har vist, at indsættelse af materialer som svamp, voks eller en brugerdefineret protese i det tomme thoracic cavity efter PNX ( dvs. proteseimplantation) svækker alveolær regenerering. Det er nu klart fastslået, at mekanisk kraft fungerer som en af ​​de vigtigste faktorer for initiering af alveolær regenerering 1 , 16 , 17 . Sådanne undersøgelser har fremhævet effektiviteten af ​​at anvende ISA-værdier fra PNX-behandlede og protese-implanterede lunger som et kriterium for kvantitativt at evaluere alveolær regenerering. Observer-bias er kendt for at påvirke målt va væsentligtLues for lungemorfologiske parametre ( fx MIL og ILV). Standardiserede protokoller kan bruges til at undgå denne bias ved bestemmelse af både ILV og MLI, som er de to parametre, der anvendes til beregning af ISA. Her giver vi meget detaljerede, standardiserede protokoller til måling af disse lungeparametre. Det er vigtigt, at evnen til nøjagtigt at kvantificere ISA løfter at forbedre pålideligheden og reproducerbarheden af ​​undersøgelser af lungefunktion i skadefremkaldte alveolære regenereringsmodeller og bør lette mekaniske opdagelser i flere lungesygdomme.

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			<td>15012-12</td>
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			<td>Fine Science Tools</td>
			<td>11000-12</td>
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			<td>12060-01</td>
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			<td>Jinhuan Medical Product Co., LTD.</td>
			<td>F602</td>
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			<td height="21" style="height:21px;">Mouse intubation platform</td>
			<td>Penn-Century, Inc</td>
			<td>Model MIP</td>
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			<td>Penn-Century, Inc</td>
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			<td height="21" style="height:21px;">Heparin sodium salt</td>
			<td>Sigma</td>
			<td>H3393</td>
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			<td height="21" style="height:21px;">Hematoxylin Solution</td>
			<td>Sigma</td>
			<td>GHS132</td>
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			<td height="21" style="height:21px;">Eosin Y solution, alcoholic</td>
			<td>Sigma</td>
			<td>HT110116</td>
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			<td height="21" style="height:21px;">10 ml Pipette</td>
			<td>Thermo Scientific</td>
			<td>170356</td>
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			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>Sigma</td>
			<td>P6148</td>
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			<td height="21" style="height:21px;">O.C.T Compound</td>
			<td>Tissue-Tek</td>
			<td>4583</td>
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			<td>Leica</td>
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			<td>Fisher HealthCare</td>
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			<td>Becton Dickinson</td>
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			<td height="21" style="height:21px;">Povidone iodine</td>
			<td>Fisher Scientific</td>
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</table>

a standardized method for measuring internal lung surface area via mouse pneumonectomy and prosthesis implantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), representing the gas-exchange capacity of the lung, is a critical criterion to assess respiratory function. However, observer bias can significantly influence measured values for lung morphological parameters. The protocol that we describe here minimizes variations during measurements of two morphological parameters used for ISA calculation: internal lung volume (ILV) and mean linear intercept (MLI). Using ISA as a morphometric and functional parameter to determine the outcome of alveolar regeneration in both pneumonectomy (PNX) and prosthesis implantation mouse models, we found that the increased ISA following PNX treatment was significantly blocked by implantation of a prosthesis into the thoracic cavity1. The ability to accurately quantify ISA is not only expected to improve the reliability and reproducibility of lung function studies in injured-induced alveolar regeneration models, but also to promote mechanistic discoveries of multiple pulmonary diseases.

<html> Vi gennemførte her et eksperiment med en PNX-behandlet gruppe og en proteseimplantation (Prothese-implanteret) gruppe. Disse grupperinger er de samme som grupperinger anvendt i en tidligere offentliggjort undersøgelse fra vores forskningsgruppe 14 . PNX- og proteseimplantationsprocedurerne er vist i figur 2 . 8 uger gamle CD-1 hanmus anvendes til operationerne og ti...

Watch this Scientific Journal Video about A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation at JoVE.com

A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation

Det indre lungeoverfladeareal (ISA) er et kritisk kriterium til vurdering af lungemorfologi og fysiologi i lungesygdomme og skadefremkaldt alveolær regenerering. Her beskrives en standardiseret metode, der kan minimere måleforspændingen for ISA i både lungepneumonektomi og protesimplantationsmusemodeller.

En standardiseret metode til måling af det indre lungeoverfladeområde via muspneumonektomi og proteseimplantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), ...

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During chronic liver injuries, progenitor cells expand in a process called ductular reaction, which also entails the appearance of inflammatory cellular ...

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts

Vurdering cardiomyocyte undertyper Efter Transcription Factor-medieret Omprogrammering af Mouse Embryonale fibroblaster

Direct reprogramming of one cell type into another has recently emerged as a powerful paradigm for regenerative medicine, disease modeling, and lineage ...

A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry

En standardiseret tilgang til multispecies Rensning af mammale mannlige germeller ved hjælp af mekanisk vævsdissociation og flowcytometri

Fluorescence-activated cell sorting (FACS) has been one of the methods of choice to isolate enriched populations of mammalian testicular germ cells. ...

A Method for Characterizing Embryogenesis in Arabidopsis

A Method for Characterizing Embryogenesis in Arabidopsis

En metode til kendetegner embryogenese i Arabidopsis

Given the highly predictable nature of their development, Arabidopsis embryos have been used as a model for studies of morphogenesis in plants. However, ...

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse

En roman brug af tre-dimensionelle høj frekvens ultralyd for tidlig graviditet karakterisering i mus

High-frequency ultrasonography (HFUS) is a common method to non-invasively monitor the real-time development of the human fetus in utero. The mouse is ...

Application of Chronic Stimulation to Study Contractile Activity-induced Rat Skeletal Muscle Phenotypic Adaptations

Anvendelse af kronisk stimulering til at studere kontraktile aktivitet-induceret rotte skeletmuskulatur fænotypiske tilpasninger

Skeletal muscle is a highly adaptable tissue, as its biochemical and physiological properties are greatly altered in response to chronic exercise. To ...

Partial Lobular Hepatectomy: A Surgical Model for Morphologic Liver Regeneration

Delvis dråbeformede Hepatectomy: En kirurgisk Model for morfologiske lever regenerering

Morphological organ regeneration following acute tissue loss is common among lower vertebrates, but is rarely observed in mammalian postnatal life. Adult ...