Amniotic Fluid Cell Culture

  • Holger Hoehn


This chapter is intended as a supplement and an update, rather than a replacement, of the chapter on this subject in the previous edition of the work (Milunsky, 1979), and the reader is encouraged to peruse that chapter along with the present one. Much valuable information deriving from the extensive personal experience of the author of that chapter (Milunsky, 1973, 1979) has remained accurate and timely, and thus will not be repeated here. This also holds for the comprehensive list of references to the early work in the field. The present chapter will deal mainly with areas in which significant recent progress has been made. One substantial advance concerns the identification and classification of colony-forming amniotic fluid cell types. Another relates to the profound impact of culture medium supplements on the success and speed of amniotic fluid cell growth.


Amniotic Fluid Prenatal Diagnosis Cloning Efficiency Amniotic Fluid Cell Chromosomal Mosaicism 
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  1. Agorastos, T., Bär, T., Grussendorf, E. I., et al., 1982, Zur Feinmorphologie der Fruchtwasserzellen, Z. Geburtshilfe Perinatol 186:41.PubMedGoogle Scholar
  2. Barnes, D., and Sato, G., 1980, Serum-free cell culture: a unifying approach, Cell 22:649.PubMedCrossRefGoogle Scholar
  3. Bergström, S., 1979, Ultrastructure of cell detachment from the human fetus in early pregnancy, Acta Obstet. Gynecol. Scand. 58:353.PubMedCrossRefGoogle Scholar
  4. Boué, J., Nicolas, H., Barichard, F., et al., 1979, Le clonage des cellules du liquide amniotique aide dans l’interprétation des mosaiques chromosomiques en diagnostic prénatal, Ann. Genet. 22:3.Google Scholar
  5. Brackertz, M., Kubbies, M., Feige, A., et al., 1983, Decreased oxygen supply enhances growth in culture of human mid-trimester amniotic fluid cells, Hum. Genet. 64:334.PubMedCrossRefGoogle Scholar
  6. Bryant, E. M., Crouch, E., Bornstein, P., et al., 1978. Regulation of growth and gene activity in euploid hybrids between human neonatal fibroblasts and epithelioid amniotic fluid cells, Am. J. Hum. Genet. 30:392.PubMedGoogle Scholar
  7. Burton, B. K., Gerbie, A. B., and Nadler, H. L., 1979, Biochemical and biological problems and pitfalls of cell culture for prenatal diagnosis, in: Genetic Disorders and the Fetus (A. Milunsky, ed.), p. 369, Plenum Press, New York.Google Scholar
  8. Chang, H. C., and Jones, O. W., 1982, Amniocentesis: Cell culture of human amniotic fluid in a hormone supplement, in: Cold Spring Harbor Conferences on Cell Proliferation, Vol. 9, p. 1187, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.Google Scholar
  9. Chang, H. C., Jones, O. W., Bradshaw, C., et al., 1981, Enhancement of human amniotic cell growth by Ficoll-Paque gradient fractionation, In vitro 17:81.PubMedCrossRefGoogle Scholar
  10. Chang, H. C., Jones, O. W., and Masui, H., 1982, Human amniotic fluid cells grown in a hormone-supplemented medium: Suitability for prenatal diagnosis, Proc. Natl. Acad. Sci. USA 79:4795.PubMedCrossRefGoogle Scholar
  11. Chen, W. W., 1982, Studies on the origin of human amniotic fluid cells by immunofluorescent staining of keratin filaments, J. Med. Genet. 19:433.PubMedCrossRefGoogle Scholar
  12. Claussen, U., 1980, The pipette method: A new rapid technique for chromosome analysis in prenatal diagnosis, Hum. Genet. 54:277.PubMedCrossRefGoogle Scholar
  13. Claussen, U., Schäfer, H., and Trampisch, H., 1984, Exclusion of chromosomal mosaicism in prenatal diagnosis, Hum. Genet. 67:23.PubMedCrossRefGoogle Scholar
  14. Cox, D. M., Niewczas-Late, V., Riffell, M. L, et al., 1974, Chromosomal mosaicism in diagnostic amniotic fluid cell cultures, Pediatr. Res. 8:679–683.PubMedCrossRefGoogle Scholar
  15. Cremer, M., Treiss, I., Cremer, T., et al., 1981, Characterization of cells of amniotic fluids by immunological identification of intermediate-sized filaments: Presence of cells of different tissue origin, Hum. Genet. 59:373.PubMedCrossRefGoogle Scholar
  16. Crickard, K., and Golbus, M. S., 1982, Influence of extracellular matrix on the proliferation of human amniotic fluid cells in vitro, Prenat. Diagn. 2:89.PubMedCrossRefGoogle Scholar
  17. Crouch, E., and Bornstein, P., 1978, Collagen synthesis by human amniotic fluid cells in culture: Characterization of a procollagen with three identical pro a (I) chains, Biochemistry 17:5495.CrossRefGoogle Scholar
  18. Crouch, E., and Bornstein, P., 1979, Characterization of a type IV procollagen synthesized by human amniotic fluid cells in culture, J. Biol. Chem. 254:4197.PubMedGoogle Scholar
  19. Crouch, E. D., Balian, G., Holbrook, K., et al., 1978, Amniotic fluid fibronectin. Characterization and synthesis by cells in culture, J. Cell Biol. 78:701.PubMedCrossRefGoogle Scholar
  20. Darlington, G. J., 1982, Application of cell-fusion techniques to induce amniotic fluid cells to express special functions and for complementation analysis, Meth. Cell Biol. 26:297.CrossRefGoogle Scholar
  21. Epstein, C. J., 1982, The use of growth factors to stimulate the proliferation of amniotic fluid cells, Meth. Cell Biol. 26:264.Google Scholar
  22. Felix, J. S., and Doherty, R. A., 1979, Amniotic fluid cell culture II. Evaluation of a red blood cell lysis procedure for culture of cells from blood-contaminated amniotic fluid, Clin. Genet. 15:215.PubMedCrossRefGoogle Scholar
  23. Felix, J. S., Doherty, R. A., Davis, H. T., et al., 1974, Amniotic fluid cell culture. I. Experimental design for evaluating cell culture variables: Determination of optimal fetal calf serum concentration, Pediatr. Res. 8:870.PubMedCrossRefGoogle Scholar
  24. Felix, J., Sun, T. T., and Littlefield, J. W., 1980, Human epithelial cells cultured from urine: Growth properties and keratin staining, In Vitro 16:866.PubMedCrossRefGoogle Scholar
  25. Franke, W. W., Appelhans, B., Schmid, E., et al., 1979. Identification and characterization of epithelial cells in mammalian tissues by immunofluorescence microscopy using antibodies to prekeratin, Differentiation 15:7.PubMedCrossRefGoogle Scholar
  26. Gerbie, A. B., Melancon, S. B., Ryan, C., et al., 1972, Cultivated epithelial-like cells and fibroblasts from amniotic fluid: Their relationship to enzymatic and cytologic analysis, Am. J. Obstet. Gynecol. 114:314.PubMedGoogle Scholar
  27. Gosden, C. M., 1983, Amniotic fluid cell types and culture. Br. Med. Bull. 39:348.PubMedGoogle Scholar
  28. Gosden, C. M., and Brock, D. J. H., 1977, Morphology of rapidly adhering amniotic fluid cells as an aid to the diagnosis of neural tube defects, Lancet 1:919.PubMedCrossRefGoogle Scholar
  29. Gosden, C.M., and Brock, D. J. H., 1978, Amniotic fluid cell morphology in early prenatal prediction of abortion and low birth weight, Br. Med. J. 2:1186.PubMedCrossRefGoogle Scholar
  30. Gospodarowicz, D., Moran, J. S., and Owashi, N. D., 1977, Effects of fibroblast growth factor and epidermal growth factor on the rate of growth of amniotic fluid-derived cells, J. Clin. Endocrinol. Metab. 44:651.PubMedCrossRefGoogle Scholar
  31. Harris, A., 1982, Glycoproteins that distinguish different cell types found in amniotic fluid, Hum. Genet. 62:188.PubMedCrossRefGoogle Scholar
  32. Harrison, H., Martin, A. O., Gemme, M. A., et al., 1983, Analysis of amniotic fluid supernatant and cellular proteins with high resolution two-dimensional electrophoresis, Am. J. Hum. Genet. 35:45A.Google Scholar
  33. Hecht, F., Peakman, D. C., Kaiser-McCaw, B., et al., 1981, Amniocyte clones for prenatal cytogenetics, Am. J. Med. Genet. 10:51.PubMedCrossRefGoogle Scholar
  34. Held, K. R., and Sönnichsen, S., 1984, The effect of oxygen tension on colony formation and cell proliferation of amniotic fluid cells in vitro, Prenat. Diagn. 4:171.PubMedCrossRefGoogle Scholar
  35. Hoehn, H., and Salk, D., 1982, Morphological and biochemical heterogeneity of amniotic fluid cells in culture, Meth. Cell Biol. 26:11.CrossRefGoogle Scholar
  36. Hoehn, H., Bryant, D. M., Karp, L. E., et al., 1974, Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. I. Clonal morphology and growth potential, Pediatr. Res. 8:746.PubMedCrossRefGoogle Scholar
  37. Hoehn, H., Bryant, E. M., Karp, L. E., et al., 1975a, Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. II. Cytogenetic parameters as functions of clonal type and preparative technique, Clin. Genet. 7:29.PubMedCrossRefGoogle Scholar
  38. Hoehn, H., Bryant, E. M., Fantel, A. G., et al., 1975b, Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. III. The fetal urine as a potential source of clonable cells, Hum. Genet. 29:285.CrossRefGoogle Scholar
  39. Hoehn, H., Rodriguez, M. L., Norwood, T. H., et al., 1978, Mosaicism in amniotic fluid cell cultures: Classification and significance, Am. J. Med. Genet. 2:253.PubMedCrossRefGoogle Scholar
  40. Holbrook, K. A., and Odland, G. F., 1975, The fine structure of developing epidermis: light, scanning, and transmission electron microscopy of the periderm. J. Invest. Dermatol. 65:16–38.PubMedCrossRefGoogle Scholar
  41. Holbrook, K. A., and Odland, G. F., 1980, Regional development of the human epidermis in the first trimester embryo and the second trimester fetus (ages related to the timing of amniocentesis and fetal biopsy). J. Invest. Dermatol. 80:161–168.CrossRefGoogle Scholar
  42. Huisjes, H. J., 1978, Cytology of the amniotic fluid and its clinical applications, in: Amniotic Fluid, Research and Clinical Application (D. V. I. Fairweather and T. K. A. B. Eskes, eds.), 2nd ed., p. 93, Elsevier/North-Holland, Amsterdam.Google Scholar
  43. Johnston, P., Salk, D., Martin, G. M., et al., 1982, Cultivated cells from mid-trimester amniotic fluids. IV. Cell type identification via one and two-dimensional electrophoresis of clonal whole cell homogenates, Prenat. Diagn. 2:79.PubMedCrossRefGoogle Scholar
  44. Kaback, M. M., Leonard, C. O., and Parmley, T. H., 1971, Intrauterine diagnosis: comparative en-zymology of cells cultivated from maternal skin, fetal skin, and amniotic fluid. Pediat. Res. 5:366–371.PubMedCrossRefGoogle Scholar
  45. Kohn, G., 1981, Failure of amniotic fluid cell cultures due to syringe toxicity, Prenat. Diagn. 1:233.PubMedCrossRefGoogle Scholar
  46. Koch, H., Bettecken, T., Kubbies, M., et al., 1984, Flow cytometric analysis of small DNA content differences in heterogeneous cell populations: Human amniotic fluid cells, Cytometry 5:118.PubMedCrossRefGoogle Scholar
  47. Medina-Gomez, P., and Bard, J. B. L., 1983, Analysis of normal and abnormal amniotic fluid cells in vitro by cinemicrography, Prenat. Diagn. 3:311.PubMedCrossRefGoogle Scholar
  48. Medina-Gomez, P., and Johnston, T. H., 1982, Cell morphology in long-term cultures of normal and abnormal amniotic fluids, Hum. Genet. 60:310.PubMedCrossRefGoogle Scholar
  49. Megaw, J. M., Priest, J. H., Priest, R. E., et al., 1977, Differentiation in human amniotic fluid cell cultures. II. Secretion of epithelial basement membrane glycoprotein, J. Med. Genet. 14:163.PubMedCrossRefGoogle Scholar
  50. Melancon, S. B., Lee, S. Y., and Nadler, H. L., 1971, Histidase activity in cultivated human amniotic fluid cells, Science 173:627.PubMedCrossRefGoogle Scholar
  51. Melnyk, J. H., Persinger, G., and Teplitz, R. L. A., 1979, A micromethod for processing amniotic fluid cells, In Vitro 15:200.Google Scholar
  52. Milunsky, A., 1973, The Prenatal Diagnosis of Hereditary Disorders, Thomas, Springfield, Illinois.Google Scholar
  53. Milunsky, A., 1979. Amniotic fluid cell culture, in: Genetic Disorders and the Fetus (A. Milunsky, ed.), p. 75, Plenum Press, New York.Google Scholar
  54. Milunsky, A., and Bender, C.S., 1979, Failure of amniotic fluid cell growth with toxic tubes, N. Engl. J. Med. 301:47.PubMedGoogle Scholar
  55. Miny, P., and Pawlowitzki, I. H., 1984, Trisomy 20 mosaicism, Prenat. Diagn. 4:411.PubMedCrossRefGoogle Scholar
  56. Moll, R., Franke, W. W., Schiller, D. L., et al., 1982, The catalogue of human cyto-keratins: Patterns of expression in normal epithelia, tumors and cultured cells, Cell 31:11.PubMedCrossRefGoogle Scholar
  57. Ochs, B. A., Franke, W. W., Moll, R., et al., 1983, Epithelial character and morphologic diversity of cell cultures from human amniotic fluids examined by immunofluorescence microscopy and gel electrophoresis of cytoskeletal proteins, Differentiation 24:153.PubMedCrossRefGoogle Scholar
  58. Osborn, M., and Weber, K., 1983, Biology of disease. Tumor diagnosis by intermediate filament typing: A novel tool for surgical pathology, Lab. Invest. 48:372.PubMedGoogle Scholar
  59. Porreco, R. P., Bradshaw, C., Sarkar, S., et al., 1980, Enhanced growth of amniotic fluid cells in presence of fibroblast growth factor, Obstet. Gynecol. 55:55.PubMedGoogle Scholar
  60. Priest, R. E., Priest, J. H., Moinuddin, J. F., et al., 1977, Differentiation in human amniotic fluid cell cultures. I. Collagen production, J. Med. Genet. 14:157.PubMedCrossRefGoogle Scholar
  61. Priest, R. E., Marimuthu, K. M., and Priest, J. H., 1978, Origin of cells in human amniotic fluid cultures. Ultrastructural features, Lab. Invest. 39:106.PubMedGoogle Scholar
  62. Priest, R. E., Priest, J. H., Moinuddin, J. F., et al., 1979, Differentiation in human amniotic fluid cell cultures: Chorionic gonadotrophin production, In Vitro 15:142.PubMedCrossRefGoogle Scholar
  63. Priest, R. E., Priest, G. H., Laundon, C. H., et al., 1980, Multinucleate cells in cultures of human amniotic fluid form by fusion, Lab. Invest. 43:140.PubMedGoogle Scholar
  64. Regauer, S., Franke, W. W., and Virtanen, I., 1985, Intermediate filament cytoskeleton of amnion epithelium and cultured amnion epithelial cells: Expression of epidermal cytokeratins in cells of a simple epithelium, J. Cell Biol. 100:997.PubMedCrossRefGoogle Scholar
  65. Salk, D. S., Disteche, C., Stenchever, M. R., et al., 1983, Routine use of Chang medium for prenatal diagnosis: Improved growth and increased chromosome breakage, Am. J. Hum. Genet. 35:151 A.Google Scholar
  66. Schmid, W., 1975, A technique for situ karyotyping of primary amniotic fluid cell cultures, Humangenetik 30:325.PubMedCrossRefGoogle Scholar
  67. Schrage, R., Bögelspacher, H. R., and Wurster, K. G., 1982, Amniotic fluid cells in the second trimester of pregnancy, Acta Cytol. 26:407.PubMedGoogle Scholar
  68. Seguin, L. R., and Palmer, C. G., 1983, Variables influencing growth and morphology of colonies of cells from human amniotic fluid, Prenat. Diagn. 3:107.PubMedCrossRefGoogle Scholar
  69. Sutherland, G. R., Bauld, R., and Bain, D. J., 1974, Observations on human amniotic fluid cell strains in serial culture, J. Med. Genet. 11:190.PubMedCrossRefGoogle Scholar
  70. Tabor, A., Lind, A. M., Andersen, A. M., et al., 1984, A culture vessel for amniotic fluid cells allowing faster preparation of chromosome slides, Prenat. Diagn. 4:451.PubMedCrossRefGoogle Scholar
  71. Therkelsen, A. J., 1979, Cell culture and cytogenetic techniques, in: Prenatal Diagnosis (J. D. Murken, S. Stengel-Rutkowski, and E. Schwinger, eds.) p. 258, Enke, Stuttgart.Google Scholar
  72. Tyden, O., Bergström, S., and Nilsson, B. A., 1981, Origin of amniotic fluid cells in mid-trimester pregnancies, Br. J. Obstet. Gynaecol. 88:278.PubMedCrossRefGoogle Scholar
  73. Uhlendorf, B.W., 1970, Use of amniotic fluid, and reliability of diagnostic procedures, in: Early diagnosis of human genetic defects. Fogarty Int. Proc. 6:149–162.Google Scholar
  74. Van der Veer, E., Kleijer, W. J., deJosselin deJong, J. E., et al., 1978, Lysosomal enzyme activities in different types of amniotic fluid cells measured by microchemical methods, combined with interference microscopy, Hum. Genet. 40:285.PubMedCrossRefGoogle Scholar
  75. Virtanen, I., von Koskull, H., Lehto, V. P., et al., 1981, Cultured amniotic fluid cells characterized with antibodies against intermediate filaments in indirect immunofluorescence microscopy, J. Clin. Invest. 68:1348.PubMedCrossRefGoogle Scholar
  76. Vlodavsky, I., Voss, R., Yarkoni, S., et al., 1982, Stimulation of human amniotic fluid cell proliferation and colony formation by cell plating on a naturally produced extracellular matrix, Prenat. Diagn. 2:13.CrossRefGoogle Scholar
  77. Von Koskull, H., Aula, P., Trejdosiewicz, L. K., et al., 1984, Identification of cells from fetal bladder epithelium in human amniotic fluid, Hum. Genet. 65:262.CrossRefGoogle Scholar

Copyright information

© Aubrey Milunsky 1986

Authors and Affiliations

  • Holger Hoehn
    • 1
  1. 1.Department of Human GeneticsUniversity of Würzburg87 WürzburgFederal Republic of Germany

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