Cytokine quantification

UMBILICAL CORD AND PLACENTAL BLOOD COLLECTION AND PROCESSING

1. Clean placental annexes using saline 0,9%.
2. Using a 10 ml syringe with needle, puncture the veins and/or arteries of the cord (Figure 1).

3. Store the blood in a heparinized tube and homogenize slowly.

    

Figure 1 - Umbilical cord blood collection.

Photo: Rodrigo Medeiros de Souza

4. To collect placental blood, make an incision in the maternal cotyledons of the placenta (Figure 2).

5. Collect 5 to 10 ml of blood of the incision, using a Pasteur pipette. Transfer the blood to a heparinized tube and homogenize slowly (Figure 2).

Figure 2 - Collection of placental blood from the maternal face of the placenta.

Photo: Rodrigo Medeiros de Souza.

6. To obtain plasma, centrifuge the heparinized blood at 3000 rpm for 10 min at RT (DO NOT USE CENTRIFUGE BRAKING - Decrease speed to 0, turn off the centrifuge and wait for the complete stop).

7. Remove the plasma with a Pasteur pipette and divide it into aliquots (ATTENTION not to aspirate the erythrocyte and leukocyte sediment).

8. Store at -20 or -80ºC until cytokine dosages.

CYTOKINE QUANTIFICATION

1. Follow the standard protocol described in the kit: BD ™ CBA Human Inflammatory Cytokines Kit. - https://www.bdbiosciences.com/ds/pm/others/551811_Book_Website.pdf.

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