Chromosomal studies on 2 mL of celomic fluid obtained during the fifth week of development in the timed-pregnant baboon model.
OBJECTIVE
To determine if chromosomal studies could be performed using 2 mL of celomicfluid obtained during the fifth postfertilization week in pregnant baboons.


METHODS
Nine ultrasound-guided celocenteses were performed. The initial 0.5 mL of celomic fluid was discarded to decrease maternal cell contamination. Approximately 2 mL of celomic fluid was then collected. The fluid was centrifuged and the supernatant removed to a final volume of 0.5 mL. The celomic fluid sample was placed in either a small plastic flaskette chamber slide with a mix of 0.5 mL celomic fluid, 1 mL of Amniomax, and 1 mL of usedfibroblast culture medium to spread on the entire surface (n=4), or a 3.5 x 1-cm plastic Petri dish with a 24 x 30-mm glass coverslip to keep the 0.5 mL celomic fluid mixed with 1 mL of Amniomax (Invitrogen, Carlsbad, California) within a 1 cm2 area (n=5). The medium was changed on day 5 and thereafter every second to third day. The cells were harvested when the number of cells appeared sufficient for chromosomal analysis.


RESULTS
Standard chromosomal studies were possible in 5 of the 9 celomicfluid samples. Mean (+/-SD) celomic fluid volume used for culture was 1.85 +/- 0.3 mL. Mean (+/-SD) time to karyotype result was 18.8 +/- 1.8 days.


CONCLUSIONS
The findings of this study suggest that there are living cells at 36-42 days of embryonic development in the extraembryonic celomic fluid of primates and that they can be cultured for chromosomal studies. However, significant improvements in understanding the biology of cells present at 5 weeks after fertilization in celomic fluid are needed to improve culture conditions.
Santolaya-Forgas
Joaquin
J
Perinatology Research Branch, National Institute of Child Health and Development, National Institutes of Health, Department of Health and Human Services, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. jsantol@med.wayne.edu
De Leon-Luis
Juan
J
Shen
Zhion
Z
McCorquodale
Maureen
M
D000818:Animals; D001826:Body Fluids / Q000166:cytology; D025063:Chromosome Disorders / Q000175:diagnosis*; D005321:Extraembryonic Membranes / Q000166:cytology*; D005240:Feasibility Studies; D005260:Female; D023421:Models, Animal; D010215:Papio; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*