TITLE: 603.3 &amp; 604.5_URMC_HTC_Lung and Lobe Processing for SenNet
AUTHORS: Gloria S Pryhuber, Heidie Huyck

[DESCRIPTION]
Purpose and Scope of the Procedure
Standardize process for processing lung donations into components for storage and distribution
Scope:       coordination of receipt and gross dissection of tissue

[GUIDELINES]
Scientific Principles 
Rapid, standardized and safe processing of human tissue for the HTC requires coordination of a team of staff, materials and attention to protocols
Rapid processing is required to maintain the tissues in a state as close to normal as possible
Ultra-High Resolution CT Scan in an inflated state will provide a high-level comparative assessment of human lung structure across developmental ages

[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. 
SECTION: 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.

SECTION: Unpacking
4. Determine BRINDL PID to assign to tissues and images by clicking the “arrived” button in BRINDL Screening log


SECTION: Unpacking
5. Record Information on Shipping Labels in Database or worksheet for entry ASAP
    i.UNOS #; Referring Company #

    ii.Courier and Tracking Numbers
SECTION: Unpacking
6. Remove Shipping Labels from package; place aside to be added to Sample File


SECTION: Unpacking
7. Open Shipping Box and Remove Styrofoam Cover

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

SECTION: Unpacking
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
        a.Is tissue submerged (usually in plastic bag, inside a hard plastic container)
        b.Are Trachea and both lungs included and intact
        c.Is Trachea Occluded, if so how
        d.Are Thymus, Spleen, Blood Sample, Lymph Nodes included?
SECTION: Grossing of Tissues
9. Maintain tissues in transplant buffer on ice. Prevent open Trachea from being submerged , +/- ETT removal
SECTION: Grossing of Tissues
10. Weigh intact organ as sent; Continue to Record all measurements and procedures in Worksheet or directly in BRINDL database                                              
SECTION: Grossing of Tissues
11. Determine displacement volume without inflation                                              
SECTION: Grossing of Tissues
12. Determine plan for organ lobes ie which to be inflated or infused and which to be dissociated                                              
SECTION: Grossing of Tissues
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)  
        a.     Weigh each tissue collected  
        b.     The non-lung samples are sectioned into approximately 0.5-1 cm3 portions  
        c.     Divide these tissues between                                                                                                      
             i.    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  
        d.     Record these tissues and how processed in BRINDL database  
SECTION: Grossing of Tissues
14.  Dissect free the bronchial branches, identify pulmonary arteries and veins   
SECTION: Grossing of Tissues
15. Divide airways and vessels to obtain 5 independent lung lobes  
SECTION: Grossing of Tissues
15.1. Weigh each lobe once isolated 
SECTION: Grossing of Tissues
15.2. Proceed with processing for each lobe as using appropriate Protocols
SECTION: Grossing of Tissues
16. Prepare accompanying tissue such as thymus, spleen, esophagus, blood, heart, in similar fashion with specific as outlined in the appropriate Protocol / SOP.
SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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SECTION: Lobe Sectioning Diagrams
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