603.3 & 604.5_URMC_HTC_Lung and Lobe Processing for SenNet

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

Record details of procedures in Worksheets.

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.

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

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

i.UNOS \#; Referring Company \#



ii.Courier and Tracking Numbers

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

Open Shipping Box and Remove Styrofoam Cover

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

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?

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

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

Determine displacement volume without inflation

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

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  

Dissect free the bronchial branches, identify pulmonary arteries and veins

Divide airways and vessels to obtain 5 independent lung lobes

Weigh each lobe once isolated

Proceed with processing for each lobe as using appropriate Protocols

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