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J. T. DINGLE AND M. WEBB
methods for the isolation and purification of these complexes, none of
which are as yet applicable to the small scale of Tissue Culture. The
only information at present available on the formation and degradation
of protein polysaccharide complexes in culture have come from the
results obtained in studies of the action of enzymes on these systems,
and will be discussed in the appropriate sections.
B. PRODUCTION OF MUCOPOLYSACCHARIDES IN CULTURE
For the purposes of this review Tissue Culture techniques may be
conveniently divided into the three main sections, tissue culture, cell
culture and organ culture. With the first of these methods the cell
population is likely to be mixed and its density not easily controlled.
These disadvantages are overcome in the second method. Isolation
of free cells by trypsin treatment, however, eliminates not only the acid
mucopolysaccharides of the intercellular material—a factor that may
be of significance if, as the work of Adams (1959a, b, 1960) suggests,
a primer or template is necessary for the synthesis of certain mucopolysaccharides but, also, it probably removes certain mucopolysaccharide
components of the cell surface. The latter effect is discussed more
completely in a subsequent section.
The media used in these three techniques vary widely, but may be
roughly divided into "natural", i.e. the serum—Tyrode—embryoextract mixture, and "synthetic" in which all the components of the
medium are chemically defined. A variant of this latter medium, which
is used frequently for the investigation of mucopolysaccharide production, is a synthetic medium that contains about 5 or 10% serum
or a protein fraction.
1. Mucopolysaccharide Production by Tissue Explants
The explant method was used in the first experiments in which the
production of mucopolysaccharides in culture was established. Thus,
Vaubel (1933) cultured synovial membrane and reported that the
"synovioblasts" were responsible for the production of synovial mucin.
The main acid mucopolysaccharide produced by cultures of human
synovial membrane was identified as hyaluronic acid by Hedberg and
Moritz (1958). This was synthesized by the peri-articular tissues as
well as by the synovial cells. Kling, Levine and Wyse (1955), Grossfeld
et al. (1955) and Gaines (1960) have also reported mucopolysaccharide
production in tissue cultures of human and mammalian synovial
membrane. Thus, it was established that explants of human and animal
synovia can form hyaluronic acid in vitro. This finding has been confirmed by a study of the synthesis of acid mucopolysaccharide by isol-
J. T. DINGLE AND M. WEBB
methods for the isolation and purification of these complexes, none of
which are as yet applicable to the small scale of Tissue Culture. The
only information at present available on the formation and degradation
of protein polysaccharide complexes in culture have come from the
results obtained in studies of the action of enzymes on these systems,
and will be discussed in the appropriate sections.
B. PRODUCTION OF MUCOPOLYSACCHARIDES IN CULTURE
For the purposes of this review Tissue Culture techniques may be
conveniently divided into the three main sections, tissue culture, cell
culture and organ culture. With the first of these methods the cell
population is likely to be mixed and its density not easily controlled.
These disadvantages are overcome in the second method. Isolation
of free cells by trypsin treatment, however, eliminates not only the acid
mucopolysaccharides of the intercellular material—a factor that may
be of significance if, as the work of Adams (1959a, b, 1960) suggests,
a primer or template is necessary for the synthesis of certain mucopolysaccharides but, also, it probably removes certain mucopolysaccharide
components of the cell surface. The latter effect is discussed more
completely in a subsequent section.
The media used in these three techniques vary widely, but may be
roughly divided into "natural", i.e. the serum—Tyrode—embryoextract mixture, and "synthetic" in which all the components of the
medium are chemically defined. A variant of this latter medium, which
is used frequently for the investigation of mucopolysaccharide production, is a synthetic medium that contains about 5 or 10% serum
or a protein fraction.
1. Mucopolysaccharide Production by Tissue Explants
The explant method was used in the first experiments in which the
production of mucopolysaccharides in culture was established. Thus,
Vaubel (1933) cultured synovial membrane and reported that the
"synovioblasts" were responsible for the production of synovial mucin.
The main acid mucopolysaccharide produced by cultures of human
synovial membrane was identified as hyaluronic acid by Hedberg and
Moritz (1958). This was synthesized by the peri-articular tissues as
well as by the synovial cells. Kling, Levine and Wyse (1955), Grossfeld
et al. (1955) and Gaines (1960) have also reported mucopolysaccharide
production in tissue cultures of human and mammalian synovial
membrane. Thus, it was established that explants of human and animal
synovia can form hyaluronic acid in vitro. This finding has been confirmed by a study of the synthesis of acid mucopolysaccharide by isol-