Amniotic fluid cell culture assay

Summary

Amniotic fluid contains fetal shed epithelial cells that can be cultured in vitro to test for fetal karyotype. Source: Tissue Culture and Molecular Cytology Techniques. (Beijing Publishing House)

Operation method

Amniotic fluid cell culture assay

Principle

Amniotic fluid contains epithelial cells shed by the fetus, which can be cultured in vitro to test for fetal karyotype.

Materials and Instruments

Iodine Alcohol Nutrient solution Serum Colchicine Acetic acid Methanol
Cotton swabs Cotton balls Tweezers Threading needles Culture flasks

Move

1. Extraction of amniotic fluid

Aseptically withdraw 10-20 ml of amniotic fluid and immediately inject it into a sterile centrifuge tube;
2. Centrifugal separation

Centrifuge at 1000 rpm for 10 minutes, remove supernatant and reserve 0.5 ml;
3. culture

Add 4 ml of culture medium (including 1 ml of calf serum), adjust the pH to 6.8 with _NaHCO3 and incubate at 37 ℃. After 7-10 days of incubation in a warm box, a large number of fibroblasts or epithelioid cells could be seen; add colchicine 0.02-0.8 ml (containing calf serum) to the culture medium. Add colchicine 0.02-0.8 μg/ml nutrient solution for 10-12 hours. 4;
4. The rest of the steps are the same as those for chromosome preparation in passaged cell culture.

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