Indium 111 - white blood cells

Radiopharmaceuticals

9: ¹¹¹In-INDIUM LABELLING OF WHITE BLOOD CELLS (with ¹¹¹In-tropolonate in plasma)
(MRC Cyclotron Unit, Hammersmith Hospital)

Method

1. Dispense 9 ml of ACD (Formula A) into each of two 60 ml syringes

2. Withdraw 51 ml of the patient's blood into each syringe using a 19 G (non-intermittent) butterfly

3. Dispense 2 ml of 6% w/v hydroxyethyl starch into 5 internally sterile 20 ml tubes

4. Dispense 20 ml of anticoagulated blood into each of the internally sterile 20 ml tubes containing the sedimentation agent, mix by one gentle inversion and leave for 45-60 min to sediment the red cells

5. Dispense the remaining 20 ml of blood into an internally sterile 20 ml tube not containing sedimentation agent and centrifuge at 1500 g for 10 min. The supernatant contains cell-free plasma and ACD which is used as the cell labelling medium and for suspending the radiolabelled cells for re-injection

6. After the red cells have sedimented (step 4), remove the leucocyte-rich platelet-rich plasma (LRPRP) into internally sterile 20 ml tubes

7. Centrifuge the LRPRP at 150 g for 5 min to give a pellet of leucocytes and a supernatant containing platelet-rich plasma (PRP)

8. Transfer PRP to an internally sterile 20 ml tube and centrifuge at 1500 g for 10 min to obtain cell-free plasma. This used for washing the cells after labelling

9. Resuspend the leucocytes in 1 ml of cell-free plasma from step 5

10. Add 0.1 ml of tropolone solution (0.054% w/v in HEPES-saline buffer, pH 7.6), followed by 30 MBq of ¹¹¹In-indium chloride solution to the cells

11. Incubate the cells for 5-10 min at room temperature

12. Add 5-10 ml of cell-free plasma from step 8 and centrifuge at 150 g for 5 min

13. Remove the supernatant and retain for determining the labelling efficiency

14. Resuspend the cells in 3-4 ml of cell-free plasma from step 5. Check the activity (20 MBq is the diagnostic reference level) and calculate the labelling efficiency

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	9: ¹¹¹In-INDIUM LABELLING OF WHITE BLOOD CELLS (with ¹¹¹In-tropolonate in plasma) (MRC Cyclotron Unit, Hammersmith Hospital)



	Method
1.	Dispense 9 ml of ACD (Formula A) into each of two 60 ml syringes
2.	Withdraw 51 ml of the patient's blood into each syringe using a 19 G (non-intermittent) butterfly
3.	Dispense 2 ml of 6% w/v hydroxyethyl starch into 5 internally sterile 20 ml tubes
4.	Dispense 20 ml of anticoagulated blood into each of the internally sterile 20 ml tubes containing the sedimentation agent, mix by one gentle inversion and leave for 45-60 min to sediment the red cells
5.	Dispense the remaining 20 ml of blood into an internally sterile 20 ml tube not containing sedimentation agent and centrifuge at 1500 g for 10 min. The supernatant contains cell-free plasma and ACD which is used as the cell labelling medium and for suspending the radiolabelled cells for re-injection
6.	After the red cells have sedimented (step 4), remove the leucocyte-rich platelet-rich plasma (LRPRP) into internally sterile 20 ml tubes
7.	Centrifuge the LRPRP at 150 g for 5 min to give a pellet of leucocytes and a supernatant containing platelet-rich plasma (PRP)
8.	Transfer PRP to an internally sterile 20 ml tube and centrifuge at 1500 g for 10 min to obtain cell-free plasma. This used for washing the cells after labelling
9.	Resuspend the leucocytes in 1 ml of cell-free plasma from step 5
10.	Add 0.1 ml of tropolone solution (0.054% w/v in HEPES-saline buffer, pH 7.6), followed by 30 MBq of ¹¹¹In-indium chloride solution to the cells
11.	Incubate the cells for 5-10 min at room temperature
12.	Add 5-10 ml of cell-free plasma from step 8 and centrifuge at 150 g for 5 min
13.	Remove the supernatant and retain for determining the labelling efficiency
14.	Resuspend the cells in 3-4 ml of cell-free plasma from step 5. Check the activity (20 MBq is the diagnostic reference level) and calculate the labelling efficiency


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