Reagents and materials:
1. Transport medium: Eagle basic medium (EBM), adding 300 U/ml penicillin and 300 μg/ml streptomycin, 150 μg/ml gentamicin and 1 μg/ml amphotericin B;
Clip
3. scissors;
4. Anticoagulant: CPDA-1: 122mmol/L (26g/L) sodium citrate, 0.142mol/L (25.5g/L) glucose, 15.6mmol/L (3.0g/L) citric acid and 18.3 Mmmol/L (2.2 g/L) sodium dihydrogen phosphate;
5. Betadine* or 70% ethanol;
6. No. 18 syringe needle;
7. Umbilical collection bag, 175ml, containing 24.5ml CPDA-1;
8. 50ml syringe containing 200I.U. heparin
experimental method:
1. After birth, use two clips to clamp the umbilical cord at the end of the near newborn and near the end of the placenta to close the umbilical cord;
2. Cut the umbilical cord between the two clips, one end close to the newborn and the other end cut off the placenta;
3. Cut the umbilical cord and transfer it to the laboratory in transport medium. The whole process is within 6-12h;
4. Wipe the needle insertion site with Betadine or 70% alcohol cotton, at the end of the umbilical cord near the fetus;
5. In order to collect the maximum amount of cord blood, the processing of the umbilical cord should be as small as possible;
6. Next, insert the needle of the collection bag or 50 ml syringe into the umbilical vein insertion site treated in step 2;
7. Keep the collection bag at a level below the insertion site to facilitate the flow of cord blood into the container under the force of gravity, or slowly withdraw the cord blood from the umbilical vein with a 50 ml syringe. Collect cord blood as much as possible, generally collecting 60-150ml;
8. If venous slippage occurs, re-insert the needle by disinfecting it with Betadine or 70% alcohol along the umbilical cord;
9. When blood flow stops, open the needle safety cap and push it into the locked position. Note: cord blood should be stored at room temperature, do not refrigerate;
10. If using a bag, hold the tube with the included retaining ring and place two safety knots as close as possible to the blood bag to prevent leakage. Then cut the needle and place the needle in the pointed container to discard it.
11. Gently turn the blood bag or syringe several times to mix the cord blood with the anticoagulant;
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