Filtration and recirculation of early amniotic fluid. Evaluation of cell cultures from 100 diagnostic cases

Due to the low cell concentration, cultures from early amniotic fluid specimens usually require 2–3 weeks in culture prior to karyotyping. The purpose of this study was to evaluate the culture quality of amniotic fluid cells from early pregnancy, obtained by a new filter technique. The hypothetical advantage of the technique was that the increased cell yield might reduce the culture time before karyotyping. Culture quality was assessed by the number of colonies, the percentage of colonies containing mitoses in filter and control cultures, and the culture time. The setting was a consecutive clinical trial. One hundred samples were obtained from ongoing pregnancies at 11–14 weeks of gestation (mean 12·8 weeks). By circulating a mean of 26 ml of amniotic fluid through a cell filter system leading the cell‐free fluid back to the amniotic cavity, the cell yield was increased in the sample of 7 ml corresponding to the dead space of the filter system. The culture results were compared with control cultures from 5 ml samples drawn from the same pregnancies prior to recirculation. The cultures from the first flushing of the filter system yielded 2·6 times more colonies and in total 4·2 times more colonies were found in the three cultures grown from each filter sample when compared with the control cultures. Moreover, the filter cultures showed significantly more colonies with mitoses. The mean culture time was 8·0 days for the filter cultures, from which the karyotypes were analysed. The controls would have needed more time in culture to fulfil the diagnostic criteria for karyotyping. One case of 47,XY, + 21 was found; the rest had normal karyotypes. We conclude that the filter technique improves the culture quality of early amniotic fluid samples and allows early arrest of the cultures.

[1]  J. Philip,et al.  Genetic amniocentesis at 7–14 weeks of gestation , 1992, Prenatal diagnosis.

[2]  K. Nicolaides,et al.  Amnifiltration in the first trimester: feasibility, technical aspects and cytological outcome , 1991, Ultrasound in Obstetrics and Gynecology.

[3]  K. Nicolaides,et al.  Randomized study of early amniocentesis versus chorionic villus sampling: a technical and cytogenetic comparison of 650 patients , 1991, Ultrasound in obstetrics & gynecology : the official journal of the International Society of Ultrasound in Obstetrics and Gynecology.

[4]  D. Coleman,et al.  Early amniocentesis at 11–14 weeks' gestation for the diagnosis of fetal chromosomal abnormality—a clinical evaluation , 1991, Prenatal diagnosis.

[5]  J. Philip,et al.  Amniocentesis with increased cell yield, obtained by filtration and reinjection of the amniotic fluid , 1991, Ultrasound in obstetrics & gynecology : the official journal of the International Society of Ultrasound in Obstetrics and Gynecology.

[6]  D. Horwell,et al.  Cytogenetic studies of amniotic fluid taken before the 15th week of pregnancy for earlier prenatal diagnosis: A report of 114 consecutive cases , 1991, Prenatal diagnosis.

[7]  S. Campbell Ian Donald's child comes of age , 1991, Ultrasound in obstetrics & gynecology : the official journal of the International Society of Ultrasound in Obstetrics and Gynecology.

[8]  K. Nicolaides,et al.  Why cell culture is successful after early amniocentesis. , 1991, Fetal Diagnosis and Therapy.

[9]  M. Sandstrom,et al.  Early Amniocentesis: Report of 407 Cases With Neonatal Follow-Up , 1990, Obstetrics and gynecology.

[10]  G. Gimelli,et al.  Genetic amniocentesis: 505 cases performed before the sixteenth week of gestation , 1990, Prenatal diagnosis.

[11]  C. Trujillo,et al.  Prospective study of amniocentesis performed between weeks 9 and 16 of gestation: its feasibility, risks, complications and use in early genetic prenatal diagnosis. , 1990, American journal of medical genetics.

[12]  J. Nevin,et al.  Early amniocentesis: Experience of 222 consecutive patients, 1987–1988 , 1990, Prenatal diagnosis.

[13]  L. Hughes,et al.  FAILURE OF TETRACHLORODECAOXYGEN ANION COMPLEX TO ASSIST WOUND HEALING , 1989, The Lancet.

[14]  C. Rodeck,et al.  Early amniocentesis: a cytogenetic evaluation. , 1989, BMJ.

[15]  S. Samuels,et al.  Amniocentesis before 15 weeks' gestation: outcome, risks, and technical problems. , 1987, American journal of obstetrics and gynecology.

[16]  Ann Tabor,et al.  RANDOMISED CONTROLLED TRIAL OF GENETIC AMNIOCENTESIS IN 4606 LOW-RISK WOMEN , 1986, The Lancet.

[17]  N. Nissen,et al.  Micropore filters for sterile filtration may leach toxic compounds affecting cell cultures (HL 60). , 1985, Experimental hematology.

[18]  A. Hislop,et al.  AMNIOCENTESIS AND LUNG GROWTH: AN ANIMAL EXPERIMENT WITH CLINICAL IMPLICATIONS , 1982, The Lancet.

[19]  W. Schmid Cytogenetical problems in prenatal diagnosis. , 2009, Hereditas.

[20]  E. Gerdts Letter: Cervical cancer and gonorrhoea. , 1974, Lancet.

[21]  MatildaM. Nelson AMNIOTIC FLUID VOLUMES IN EARLY PREGNANCY , 1972, The Journal of obstetrics and gynaecology of the British Commonwealth.