Embryonic Organs in Culture

  • D. M. Kochhar
Part of the Handbook of Experimental Pharmacology book series (HEP, volume 65)


It is not my purpose to review the field of experimental embryology that deals with tissue culture of mammalian embryonic organs (Wessells 1967). A number of embryonic organs and tissues are known to undergo morphogenesis and attain functional maturation upon explanation in vitro, and their capabilities have been exploited to a variable degree in investigations on developmental mechanisms and the processes of cell differentiation (Saxén and Kohonen 1969). Some examples are: kidney (Grobstein 1956; Saxén et al. 1968), sex organs (Odor and Blandau 1971; Haffen 1976), pancreas (Sarras et al. 1981; Maylde-Pfenninger and Jamieson 1981; Grobstein 1953), skin (Sugimoto and Endo 1969), tooth (Thesleff 1976; Thesleff et al. 1981), palate (Moriarty et al. 1963; Lahti et al. 1972; Newall and Edwards 1976, 1981; Greene and Pratt 1977), other craniofacial tissues (Johnston and Listgarten 1972; Johnston et al. 1977; Greenberg and Schrier 1977), and limbs (Agnish and Kochhar 1977; Kochhar et al. 1976). Any of these or other developing organs are suitable subjects for the development of a teratologic screening system. This chapter, however, is limited to just one example, i.e., limb bud, that may suffice to bring out feasibility, advantages, and shortcomings of organ culture system in general for short-term teratologic screening purposes.


Organ Culture Cytosine Arabinoside Retinyl Palmitate Organ Culture System Apical Ectodermal Ridge 
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  1. Agnish ND, Kochhar DM (1976 a) Direct exposure of post-implantation mouse embryos to 5-bromodeoxyuridine in vitro and its effects on subsequent chondrogenesis in the limbs. J Embryol Exp Morphol 36:623–638PubMedGoogle Scholar
  2. Agnish ND, Kochhar DM (1976 b) Direct exposure of mouse embryonic limb-buds to 5-brqmodeoxyuridine in vitro and its effect on chondrogenesis: increasing resistance to the analog at successive stages of development. J Embryol Exp Morphol 36:639–652PubMedGoogle Scholar
  3. Agnish ND, Kochhar (1977) The role of somites in the growth and early development of mouse limb buds. Dev Biol 56:174–183PubMedCrossRefGoogle Scholar
  4. Ames BN, Lee FD, Durston WE (1973) An improved bacterial test system for the detection and classification of mutagens and carcinogens. Proc Natl Acad Sci USA 70:782–786PubMedCrossRefGoogle Scholar
  5. Ashby R, Davis L, Dewhurst BB, Espinal R, Penn RN, Upshall DG (1976) Aspects of teratology of cyclophosphamide. Cancer Treat Rep 60:477–482PubMedGoogle Scholar
  6. Aydelotte MB, Kochhar DM (1972) Development of mouse limb buds in organ culture; chondrogenesis in the presence of proline analog, L-azetidine-2-carboxylic acid. Dev Biol 28:191–201PubMedCrossRefGoogle Scholar
  7. Aydelotte MB, Kochhar DM (1975) Influence of 6-diazo-5-oxo-L-norleucine (DON), a glutamine analogue, on cartilaginous differentiation in mouse limb buds in vitro. Differentiation 4:73–80PubMedCrossRefGoogle Scholar
  8. Barrach HJ, Bauman I, Neubert D (1978) The applicability of in vitro systems for the evaluation of the significance of pharmacokinetic parameters for the induction of an embryonic effect. In: Neubert D, Merker HJ, Nau H, Langman J (eds) Role of pharmacokinetics in prenatal and perinatal toxicology. G Thieme, Stuttgart, pp 323–336Google Scholar
  9. Caplan AI (1981) The molecular control of muscle and cartilage development. In: Subtelny S, Abbott UK (eds) Levels of genetic control in development. AR Liss, New York, pp 37–68Google Scholar
  10. Ebert JD, Marois M (1976) Tests of teratogenicity in vitro. North-Holland, AmsterdamGoogle Scholar
  11. Fell HB, Landauer W (1935) Experiments on skeletal growth and development in vitro in relation to the problem of avian phokomelia. Proc R Soc Lond [Biol] 118:133–154CrossRefGoogle Scholar
  12. Fouts JR (1973) Microsomal mixed-function oxidases in the fetal and newborn rabbit. In: Boreus L (ed) Fetal pharmacology. Raven Press, New York, pp 305–320Google Scholar
  13. Fouts JR, Adamson RH (1959) Drug metabolism in the newborn rabbit. Science 129:897–898PubMedCrossRefGoogle Scholar
  14. Gibson JE, Becker BA (1968) The teratogenicity of cyclophosphamide in mice. Cancer Res 28:475–480PubMedGoogle Scholar
  15. Greenberg JH, Schrier BK (1977) Development of choline acetyltransferase activity in chick cranial neural crest cells in culture. Dev Biol 61:86–93PubMedCrossRefGoogle Scholar
  16. Greene RM, Pratt RM (1977) Inhibition by DON of rat palatal glycoprotein synthesis and epithelial cell adhesion in vitro. Exp Cell Res 105:27–37PubMedCrossRefGoogle Scholar
  17. Grobstein C (1953) Morphogenetic interaction between embryonic mouse tissues separated by a membrane filter. Nature 172:869–871PubMedCrossRefGoogle Scholar
  18. Grobstein C (1956) Transfilter induction of tubules in mouse metanephric mesenchyme. Exp Cell Res 10:424–440PubMedCrossRefGoogle Scholar
  19. Johnston MC, Listgarten MA (1972) The migrations, interactions, and early differentiation of oro-facial tissues. In: Slavkin HC, Bavetta LA (eds) Developmental aspects of oral biology. Academic Press, New York, pp 53–80Google Scholar
  20. Johnston MC, Morriss GM, Kuchner DC, Bingle GJ (1977) Abnormal organogenesis of facial structures. In: Wilson JG, Fraser FC (eds) Handbook of teratology, vol 2. Plenum Press, New York, pp 421-451Google Scholar
  21. Haffen K (1976) Organ culture and determination of sex organs. In: Ebert JD, Marois M (eds) Tests of teratogenicity in vitro. North-Holland, Amsterdam, pp 285–301Google Scholar
  22. Hascall VC, Oegema TR, Brown M, Caplan AI (1976) Isolation and characterization of proteoglycans from chick hmb bud chondrocytes grown in vitro. J Biol Chem 251:3511–3519PubMedGoogle Scholar
  23. Hinchliffe JR, Johnson DR (1980) The development of the vertebrate limb. Clarendon Press, OxfordGoogle Scholar
  24. Kochhar DM (1970) Effects of azetidine-2-carboxylic acid, a proline analog, on chondrogenesis in cultured limb buds. In: Bass R, Beck F, Merker HJ, Neubert D, Randham B (eds) Metabolic pathways in mammalian embryos during organogenesis and its modification by drugs. Freie Universität Press, Berlin, pp 475–482Google Scholar
  25. Kochhar DM (1973) Limb development in mouse embryos. I. Analysis of teratogenic effects of retinoic acid. Teratology 7:289–298CrossRefGoogle Scholar
  26. Kochhar DM (1975) The use of in vitro procedures in teratology. Teratology 11:273–288PubMedCrossRefGoogle Scholar
  27. Kochhar DM (1977) Cellular basis of congenital limb deformity induced in mice by vitamin A. In: Bergsma D, Lenz W (eds) Morphogenesis and malformation of the limb. AR Liss, New York, pp 111–154Google Scholar
  28. Kochhar DM, Agnish ND (to be published) Teratogenesis testing in vitro. In: Nardone RM (ed) Toxicity testing in vitro. Academic Press, New YorkGoogle Scholar
  29. Kochhar DM, Aydelotte MB (1974) Susceptible stages and abnormal morphogenesis in the developing mouse limb, analyzed in organ culture after transplacental exposure to vitamin A (retinoic acid). J Embryol Exp Morphol 31:721–734PubMedGoogle Scholar
  30. Kochhar DM, Aydelotte MB, Vest TK (1976) Altered collagen fibrillogenesis in embryonic mouse limb cartilage deficient in matrix granules. Exp Cell Res 102:213–222PubMedCrossRefGoogle Scholar
  31. Kochhar DM, Penner JD, Knudsen TB (1980) Embryotoxic, teratogenic and metabolic effects of ribavirin in mice. Toxicol Appl Pharmacol 52:99–112PubMedCrossRefGoogle Scholar
  32. Kwasigroch TE, Skalko RG, Church JK (1981) Development of limb buds in organ culture: examination of hydroxyurea enhancement of bromodeoxyuridine toxicity using image analysis. In: Neubert D, Merker H-J (eds) 5th symposium on prenatal development: applicability of culture techniques for studies on prenatal differentiation and toxicity. W de Gruyter, Berlin, pp 237–253Google Scholar
  33. Lahti A, Antila E, Saxén L (1972) The effect of hydrocortisone on the closure of the palatal shelves in two inbred strains of mice in vivo and in vitro. Teratology 6:37–42PubMedCrossRefGoogle Scholar
  34. Linsenmayer TF, Kochhar DM (1979) In vitro cartilage formation: effects of 6-diazo-5-oxo-L-norleucine (DON) on glycosaminoglycan and collagen synthesis. Dev Biol 69:517–528PubMedCrossRefGoogle Scholar
  35. Manson JM, Simons R (1979) In vitro metabolism of cyclophosphamide in limb bud culture. Teratology 19:149–158PubMedCrossRefGoogle Scholar
  36. Maylie-Pfenninger M-F, Jamieson JD (1981) Effect of 5-bromodeoxyuridine on appearance of cell-surface saccharides in organ cultures of embryonic pancreas. Dev Biol 87:16–23CrossRefGoogle Scholar
  37. Miller EJ, Matukas VJ (1969) Chick cartilage collagen: a new type of α1 chain not present in bone or skin of the species. Proc Natl Acad Sci USA 64:1264–1268PubMedCrossRefGoogle Scholar
  38. Moriarty TM, Weinstein S, Gibson RD (1963) The development in vitro and in vivo of fusion of the palatal processes of rat embryos. J Embryl Exp Morphol 11:605–619Google Scholar
  39. Neubert D, Barrach HJ (1977) Significance of in vitro techniques for the evaluation of embryotoxic effects. In: Neubert D, Merker HJ, Kwasigroch TE (eds) Methods in prenatal toxicology: evaluation of embryotoxic effects in experimental animals. G Thieme, Stuttgart, pp 202–209Google Scholar
  40. Neubert D, Merker H-J, Barrach H-J, Lessmollmann U (1976) Biochemical and teratological aspects of mammalian limb bud development in vitro. In: Ebert JD, Marois M (eds) Tests of teratogenicity in vitro. North-Holland, Amsterdam, pp 335–365Google Scholar
  41. Neubert D, Hinz N, Baumann I, Barrach HJ, Schmidt K (1978 a) Attempt upon a quantitative evaluation of the degree of differentiation of or the degree of interference with development in organ culture. In: Neubert D, Merker HJ, Nau H, Langman J (eds) Role of pharmacokinetics in prenatal and perinatal toxicology. G Thieme, Stuttgart, pp 337– 349Google Scholar
  42. Neubert D, Merker HJ, Nau H, Langman J (eds) (1978 b) Role of pharmacokinetics in prenatal and perinatal toxicology. G Thieme, StuttgartGoogle Scholar
  43. Newall DR, Edward JRG (1976) In vitro fusion of the human secondary palate. Cleft Palate J 13:54–56PubMedGoogle Scholar
  44. Newall DR, Edwards JRG (1981) The effect of vitamin A on fusion of mouse palates. II.Retinyl palmitate, retinol, and retinoic acid in vitro. Teratology 23:125–130PubMedCrossRefGoogle Scholar
  45. Odor DL, Blandau RJ (1971) Organ cultures of fetal mouse ovaries. I.Light microscopic structure. Am J Anat 131:387–414PubMedCrossRefGoogle Scholar
  46. Pelkonen O, Jouppila P, Karki NT (1973) Attemps to induce drug metabolism in human fetal liver and placenta by administration of phenobarbital to mothers. Arch Int Pharmacodyn 202:288–297PubMedGoogle Scholar
  47. Rane A, Berggren M, Yaffee S, Ericsson JLE (1973) Oxidative drug metabolism in the perinatal rabbit liver and placenta. Xenobiotica 3:37–48PubMedCrossRefGoogle Scholar
  48. Sarras MP Jr, Maylie-Pfenninger M-F, Manzi RM, Jamieson JD (1981) The effects of tunicamycin on development of the mammalian embryonic pancreas. Dev Biol 87:1–15PubMedCrossRefGoogle Scholar
  49. Saunders JW Jr (1948) The proximo-distal sequence of the origin of the parts of the chick wing and the role of ectoderm. J Exp Zool 108:363–404PubMedCrossRefGoogle Scholar
  50. Saxén L, Kohonen J (1969) Inductive tissue interactions in vertebrate morphogenesis. Int Rev Exp Pathol 8:57–128PubMedGoogle Scholar
  51. Saxén L, Koskimies O, Lahti A, Miettinen H, Rapola J, Wartiovaara J (1968) Differentiation of kidney mesenchyme in an experimental model system. Adv Morphog 7:251–293PubMedGoogle Scholar
  52. Saxén L, Karkinen-Jääsklainen M, Saxen I (1976) Organ culture in teratology. In: Gropp A, Benirschke K (eds) Current topics in pathology. Springer, Berlin Heidelberg New York, pp 123–143Google Scholar
  53. Shepard TH, Bass GL (1970) Organ culture of limb buds from riboflavin-defîcient and normal rat embryos in normal and riboflavin-deficient media. Teratology 3:163–168PubMedCrossRefGoogle Scholar
  54. Solursh M, Ahrens PB, Reiter RS (1978) A tissue culture analysis of the steps in limb ehon-drogenesis. In Vitro 14:51–61PubMedCrossRefGoogle Scholar
  55. Stark RJ, Searls RL (1973) A description of chick wing bud development and a model of limb morphogenesis. Dev Biol 33:138–153PubMedCrossRefGoogle Scholar
  56. Stark RJ, Searls RL (1974) The establishment of the cartilage pattern in the embryonic chick wing and evidence for a role of the dorsal and ventral ectoderm in normal wing development. Dev Biol 38:51–63PubMedCrossRefGoogle Scholar
  57. Subtelny S, Abbott UK (1981) Levels of genetic control in development. AR Liss, New YorkGoogle Scholar
  58. Sugimoto M, Endo H (1969) Effects of hydrocortisone on the keratinization of chick embryonic skin growing in a chemically defined medium. Nature 222:1270–1272PubMedCrossRefGoogle Scholar
  59. Thesleff I (1976) Differentiation of odontogenic tissues in organ culture. Scand J Dent Res 84:353–356PubMedGoogle Scholar
  60. Thesleff I, Barrach H-J, Foidart JM, Vaheri A, Pratt RM, Martin GR (1981) Changes in distribution of type IV collagen, laminin, proteoglycan and fibronectin during mouse tooth development. Dev Biol 81:182–192PubMedCrossRefGoogle Scholar
  61. Toledo TM, Harper RC, Moser RH (1971) Fetal effects during cyclophosphamide and irradiation therapy. Ann Intern Med 74:87–91PubMedGoogle Scholar
  62. Trowell OA (1954) A modified technique for organ culture in vitro. Exp Cell Res 6:246–248PubMedCrossRefGoogle Scholar
  63. Trowell OA (1961) Problems in the maintenance of mature organs in vitro. In: La culture organotypique. Editions du Centre National de la Recherche Scientifique, Paris, pp 237–249Google Scholar
  64. von der Mark K, Conrad G (1979) Cartilage cell differentiation. Clin Orthop 139:185–205PubMedGoogle Scholar
  65. Wessells NK (1967) Avian and mammalian organ culture. In: Wilt FH, Wessells NK (eds) Methods in developmental biology. TY Crowell, New York, pp 445–456Google Scholar
  66. Yaffee SJ, Rane A, Sjogvist F, Boreus LO, Orrenius S, (1970) The presence of a monoxy-genase system in human fetal liver microsomes. Life Sci 9:1189–1200CrossRefGoogle Scholar

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© Springer-Verlag Berlin Heidelberg 1983

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  • D. M. Kochhar

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