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    603.3 & 604.5_URMC_HTC_Lung and Lobe Processing for SenNet

    Gloria S Pryhuber, Heidie Huyck

    Published: 2023-09-21 DOI: 10.17504/protocols.io.n2bvj395plk5/v1
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    Abstract

    Purpose and Scope of the Procedure

    1. Standardize process for processing lung donations into components for storage and distribution
    2. Scope: coordination of receipt and gross dissection of tissue

    Steps

    1.

    Assemble Team and Materials as needed for processing, to accomplish: i.CT Scan

    ii.Separation of Lobes
    
    iii.Tissue Dissociation
    
    iv.Formalin Inflation
    
    v.OCT /CMC Inflation
    
    vi.Paraformaldehyde Inflation
    
    2.

    Record details of procedures in Worksheets.

    Unpacking

    3.

    Shipped package is opened on arrival to remove a blood sample, if provided, as it should be kept at room temperature.

    The remainder of the package is to be kept in cold room until processing begins to maintain approximately 4°C temperature of tissue.

    4.

    Determine BRINDL PID to assign to tissues and images by clicking the “arrived” button in BRINDL Screening log

    5.

    Record Information on Shipping Labels in Database or worksheet for entry ASAP

    i.UNOS \#; Referring Company \#
    
    
    
    ii.Courier and Tracking Numbers
    
    6.

    Remove Shipping Labels from package; place aside to be added to Sample File

    7.

    Open Shipping Box and Remove Styrofoam Cover

    i.Note Condition of Packing in Database (ice sufficient, layered packaging, no leakage, etc.)

    8.

    6.Open container containing tissue and move lung tissue to a clean plastic bag on ice.

    i.Note Condition of Lung Tissue in Database
    
    1.Is tissue submerged (usually in plastic bag, inside a hard plastic container)
    
    2.Are Trachea and both lungs included and intact
    
    3.Is Trachea Occluded, if so how
    
    4.Are Thymus, Spleen, Blood Sample, Lymph Nodes included?
    

    Grossing of Tissues

    9.

    Maintain tissues in transplant buffer on ice. Prevent open Trachea from being submerged , +/- ETT removal

    10.

    Weigh intact organ as sent; Continue to Record all measurements and procedures in Worksheet or directly in BRINDL database

    11.

    Determine displacement volume without inflation

    12.

    Determine plan for organ lobes ie which to be inflated or infused and which to be dissociated

    13.

    Remove excess tissues around trachea and large airways at hilum

    Collect lymph nodes, excess large vessels, esophagus and nerves 
    
        (Best to place these back in storage buffer and Tend to Lung First)  
    
    1.     Weigh each tissue collected  
    
    2.     The non-lung samples are sectioned into approximately 0.5-1 cm3 portions  
    
    3.     Divide these tissues between                                                                                                      
    
    9.    10% formalin, place in labeled tissue cassette in formalin
    
                    Store at `4°C` to fix for 44-48 hrs, 
    
    
    
                    Section at mid-time.  
    
                    Volume of fixative to tissue should be approximately 10:1.  
    
            ii.     PFA: place in 4%PFA stored at `4°C` to fix for 20-28 hours before moving to 30%Sucrose  
    
    
    
    4.     Record these tissues and how processed in BRINDL database  
    
    14.

    Dissect free the bronchial branches, identify pulmonary arteries and veins

    15.

    Divide airways and vessels to obtain 5 independent lung lobes

    15.1.

    Weigh each lobe once isolated

    15.2.

    Proceed with processing for each lobe as using appropriate Protocols

    16.

    Prepare accompanying tissue such as thymus, spleen, esophagus, blood, heart, in similar fashion with specific as outlined in the appropriate Protocol / SOP.

    Lobe Sectioning Diagrams

    17.
    Separation into lobes at bronchi
    Separation into lobes at bronchi
    18.
    Left Lower Lobe Tissue Blocking Strategy
    Left Lower Lobe Tissue Blocking Strategy
    19.
    Right Lower Lobe Tissue Blocking Strategy
    Right Lower Lobe Tissue Blocking Strategy
    20.
    Left Upper Lobe Tissue Blocking Strategy
    Left Upper Lobe Tissue Blocking Strategy
    21.
    Right Middle Lobe Biopsy and Dissociation Strategy
    Right Middle Lobe Biopsy and Dissociation Strategy
    22.
    Right Middle Lobe Tissue Blocking Strategy - alternating slices flash frozen / FFPE / PFAFrozenOCT
    Right Middle Lobe Tissue Blocking Strategy - alternating slices flash frozen / FFPE / PFAFrozenOCT
    23.
    Trachea and Bronchi Tissue Blocking Strategy - image shows one side of bisected trachea only
    Trachea and Bronchi Tissue Blocking Strategy - image shows one side of bisected trachea only
    24.
    Spleen Tissue Blocking Strategy
    Spleen Tissue Blocking Strategy

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