. 3. C O N S T R U C T I O N A N D USE OF S Y N T H E T I C M E D I A
127
range of concentrations. Limited success, with slow logarithmic
growth of strain N C T C 2071 in synthetic medium and methylcellulose
has been achieved by Bryant et al. (1961) in vessels coated with silicone.
C. O R G A N C U L T U R E S IN S Y N T H E T I C M E D I A
Most work on the culture of organs, or organ fragments, has involved
experiments of short duration—sometimes hours only, more often a
few days, and only rarely weeks (see Chapter 2). It is to be expected,
therefore, that the nutritional needs of such cultures might be simpler
than those of cell cultures of long duration, particularly in respect of,
for example, the B vitamins, for, as was noted above, evidence of
deficiences in cell cultures may not be demonstrable for several weeks.
Wolff, Haffen, Kieny and Wolff (1953) started their work with a very
simple medium (salts, glucose and amino acids) and obtained growth
and differentiation of embryonic organs during periods up to seven
days. Addition of /?-aminobenzoic acid, but no other vitamins, stimulated the differentiation of chick and duck tibia, syrinx, testis and
ovary. They already at that time observed nutritional differences
between tissues, later substantiated (Wolff, Haffen and Dieterlen, 1960)
by the observation that a medium which is sufficient for the differentiation of ovaries needs, for differentiation of testes, supplementation
with several additional amino acids. One of the main objectives of most
organ culture has been to retain, or to demonstrate the development of,
specific functional capacities in the organs studied. The study of differentiation and function in vitro has been used as a means of investigating
the effects of particular hormones on known or probable target organs.
Media for organ culture, therefore, while they may be less nutritionally
complete than those for long-term cell culture, are more often specially
supplemented to meet particular needs.
The special requirements of functional organs in synthetic media
are well exemplified by the work of Elias (1957), Elias and Rivera
(1959) and of Rivera and Bern (1961) on mammary tissue. Using the
defined medium 199 (Morgan, Morton and Parker, 1950), supplemented with various hormones, and the Chen technique, Elias stimulated pre-lactating mammary tissue of C3H/HeCrgl mice to secretion
with a mixture of oestrone, progesterone, Cortisol and pituitary
hormones. The full activity of this mixture was associated with Cortisol
and mammotrophic hormone. Rivera and Bern (1961), using tissues
from the same strain of mouse, demonstrated that late pre-lactating,
early pre-lactating and non-pregnant glands all required insulin, and
that the requirement for Cortisol for maintenance of alveolar structure
was greater, the more differentiated the tissue. In the presence of both
these hormones, the pituitary hormones, mammotrophin and somato-
127
range of concentrations. Limited success, with slow logarithmic
growth of strain N C T C 2071 in synthetic medium and methylcellulose
has been achieved by Bryant et al. (1961) in vessels coated with silicone.
C. O R G A N C U L T U R E S IN S Y N T H E T I C M E D I A
Most work on the culture of organs, or organ fragments, has involved
experiments of short duration—sometimes hours only, more often a
few days, and only rarely weeks (see Chapter 2). It is to be expected,
therefore, that the nutritional needs of such cultures might be simpler
than those of cell cultures of long duration, particularly in respect of,
for example, the B vitamins, for, as was noted above, evidence of
deficiences in cell cultures may not be demonstrable for several weeks.
Wolff, Haffen, Kieny and Wolff (1953) started their work with a very
simple medium (salts, glucose and amino acids) and obtained growth
and differentiation of embryonic organs during periods up to seven
days. Addition of /?-aminobenzoic acid, but no other vitamins, stimulated the differentiation of chick and duck tibia, syrinx, testis and
ovary. They already at that time observed nutritional differences
between tissues, later substantiated (Wolff, Haffen and Dieterlen, 1960)
by the observation that a medium which is sufficient for the differentiation of ovaries needs, for differentiation of testes, supplementation
with several additional amino acids. One of the main objectives of most
organ culture has been to retain, or to demonstrate the development of,
specific functional capacities in the organs studied. The study of differentiation and function in vitro has been used as a means of investigating
the effects of particular hormones on known or probable target organs.
Media for organ culture, therefore, while they may be less nutritionally
complete than those for long-term cell culture, are more often specially
supplemented to meet particular needs.
The special requirements of functional organs in synthetic media
are well exemplified by the work of Elias (1957), Elias and Rivera
(1959) and of Rivera and Bern (1961) on mammary tissue. Using the
defined medium 199 (Morgan, Morton and Parker, 1950), supplemented with various hormones, and the Chen technique, Elias stimulated pre-lactating mammary tissue of C3H/HeCrgl mice to secretion
with a mixture of oestrone, progesterone, Cortisol and pituitary
hormones. The full activity of this mixture was associated with Cortisol
and mammotrophic hormone. Rivera and Bern (1961), using tissues
from the same strain of mouse, demonstrated that late pre-lactating,
early pre-lactating and non-pregnant glands all required insulin, and
that the requirement for Cortisol for maintenance of alveolar structure
was greater, the more differentiated the tissue. In the presence of both
these hormones, the pituitary hormones, mammotrophin and somato-