Table Of ContentThe Cultured Cell and
Inherited Metabolic Disease
Previous Symposia of the Society for the
Study of Inborn Errors of Metabolism-
1. Neurometabolic Disorders in Childhood. Ed. K. S. Holt and J. Milner
1963
2. Biochemical Approaches to Mental Handicap in Children. Ed. J. D.
Allan and K. S. Holt 1964
3. Basic Concepts of Inborn Errors and Defects of Steroid Biosynthesis. Ed.
K. S. Holt and D. N. Raine 1965
4. Some Recent Advances in Inborn Errors of Metabolism. Ed. K. S. Holt
and V. P. Coffey 1966
5. Some Inherited Disorders of Brain and Muscle. Ed. J. D. Allan and D.
N. Raine 1969
6. Enzymopenic Anaemias, Lysosomes and other papers. Ed. J. D. Allan, K.
S. Holt, J. T. Ireland and R. J. Pollitt 1969
7. Errors of Phenylalanine Thyroxine and Testosterone Metabolism. Ed. W.
Hamilton and F. P. Hudson 1970
8. Inherited Disorders of Sulphur Metabolism. Ed. N. A. J. Carson and D.
N. Raine 1971
9. Organic Acidurias. Ed. J. Stern and C. Toothill 1972
10. Treatment of Inborn Errors of Metabolism. Ed. J. W. T. Seakins, R. A.
Saunders and C. Tooth ill 1973
11. Inborn Errors of Skin, Hair and Connective Tissue. Ed. J. B. Holton and
J. T. Ireland 1975
12. Inborn Errors of Calcium and Bone Metabolism. Ed. H. Bickel and J.
Stern 1976
13. Medico-Social Management of Inherited Metabolic Disease. Ed. D. N.
Raine 1977
The Society exists to promote exchanges of ideas between workers in different
disciplines who are interested in any aspect of inborn metabolic disorders. Par
ticulars of the Society can be obtained from the Editors of this Symposium.
• Symposia 1-10 published by E. & S. Livingstone
The Cultured Cell and
Inherited Metabolic Disease
MONOGRAPH BASED UPON
Proceedings of the Fourteenth Symposium of
The Society for the Study of Inborn Errors of Metabolism
EDITED BY
R. A. Harkness
and
F. Cockburn
MTP
Published by
MTP Press Limited
PO Box 55, St. Leonard's House
St. Leonardgate, Lancaster, England
© 1977 The Society for the Study of Inborn Errors of Metabolism
Softcover reprint of the hardcover 1st edition 1977
First Published 1977
No part of this book may be reproduced
in any form without permission from the
publisher except for the quotation of
brief passages for the purposes of review
ISBN-I3: 978-94-011-6629-4 e-ISBN-I3: 978-94-011-6627-0
DOl: 10.1007/978-94-011-6627-0
Text set in 11/12 pt Photon Imprint,
Great Britain at The Pitman Press, Bath
Contents
Preface Vll
List of Contributors Xl
Cell Biology
1 Cell Biology and Cell Culture Methods-A Review 3
D. G. Harnden
2 Influence of Cell Culture Medium on the Metabolic Behaviour
of the Fibroblast 16
S. O. Lie
3 Density-dependent Growth Regulation in Cultures of Human
Diploid Fibroblasts 33
T. Elsdale
4 Cryopreservation of Tissue Culture Cells 4·3
D. E. Pegg
5 Tissue Culture in the Study of Neuromuscular Diseases 57
V. Dubowitz
6 Culture of Neurons and Glial Cells 69
J.
E. Thompson
Biochemistry
7 The Biochemistry of Cytodifferentiation: an Outline of Progress 79
D. E. S. Truman
8 A Comparison of Leucocytes and Cultured 'Fibroblasts' in
Diagnosis 90
R. A. Harkness
9 Acid Hydrolase Production, Release and Uptake by Cultured
Fibroblasts 105
K. von Figura
10 Sulphatase Deficiencies 120
A. B. Roy
VI THE CULTURED CELL AND INHERITED METABOLIC DISEASE
11 Clinical, Biochemical and Genetic Heterogeneity in Gangliosidoses 13
H. Galjaard and A. J. J. Reuser
12 The Cell Membrane in Metabolic Control: The Regulation of
Cholesterol Biosynthesis in Familial Hypercholesterolaemia 16
F. W. Robertson
Milner Lecture
13 Newer Developments in Tissue Culture: Further Aid in the Study
of Inborn Errors of Metabolism 18~
J. E. Seegmiller
Short Papers
Index 20
Preface
The use of cultured cells in the clinical diagnosis of hereditary metabolic dis
ease is a rapidly developing subject to which many different disciplines have
brought their expertise and knowledge. A number of scientists who have in
dividually contributed to the growth of the subject gave invited papers at the
Fourteenth Symposium of the Society for the Study of Inborn Errors of
Metabolism in the University of Edinburgh on 13-16th July, 1976. These
papers form the basis of this monograph which brings together contributions
from the basic sciences and from physicians concerned primarily with human
disease. The cross-fertilization produced by this interdisciplinary communica
tion was invaluable to those trying to understand and overcome diagnostic
problems posed by hereditary metabolic disease.
Cell culture methods and cell preservation techniques were described by D.
G. Harnden and D. E. Pegg; Dr T. Elsdale outlined some of the factors which
control in vitro cell growth and division. Cell culture methods and cryopreser
vation techniques have allowed the wide distribution of biochemically abnor
mal cells and their study over long periods of time. It is also evident that when
a defect which produces severe metabolic disorder in man can be studied in the
laboratory using isolated cell cultures a wide variety of investigative procedures
can be focused on to the cellular defect without distress or discomfort to the
patient or relatives. Many different laboratories employing their own
specialized techniques can contribute to the diagnosis, management, cure and
prevention of the metabolic defect. Confirmation of results is needed because
there are many pitfalls in work on cultured cells as shown by S. O. Lie in his
contribution on the influence of cell culture medium on the metabolic behaviour
of the fibroblast.
Cultured cells have so far been used extensively in studies on generally dis
tributed systems such as the lysosomal acid hydrolases. In recent years the
culture of specialized cells has been developing and is reviewed for
neuromuscular systems by V. Dubowitz and for neurons and glial cells by E. J.
Thompson. Both have suggested possible future applications for these sur
prisingly highly developed methods.
viii THE CULTURED CELL AND INHERITED METABOLIC DISEASE
The biochemical section opens with an outline of the biochemistry of
cytodifferentiation which shows many areas of our present knowledge with no
apparent 'pathology' and aptly concludes with a plea to clinicians for evidence
of such abnormalities. In the following comparison of leucocytes and cultured
'fibroblasts' in diagnosis, the difference is shown between a differentiated cell,
the phagocytic microbicidal polymorphonuclear neutrophil leucocyte and the
basic undifferentiated cultured 'fibroblast'. This section also contains a
framework for the individual contributions to this monograph and an outline of
present practice in the diagnosis of hereditary metabolic disease.
The next three contributions are concerned with deficiencies of lysosomal
acid hydrolases which cause a wide range of storage disorders. The value of
communication between the basic sciences and medicine is beautifully il
lustrated by these papers from Germany, Holland and Australia.
Defects in control of biosynthesis is considered next with a review of a defect
in the control of cholesterol biosynthesis by a geneticist, F. W. Robertson.
Finally in the Milner Lecture, Newer Developments in Tissue Culture:
Further Aid in the Study of Inborn Errors of Metabolism, Professor J. E.
Seegmiller's own many contributions to our knowledge of the biochemistry of
purines are reviewed. He also discusses the transformation of lymphocytes and
the establishment of stable cell lines which carry defects. This exciting develop
ment may well further the progress of the study of inborn errors of metabolism,
and of biochemistry. It should also help the many families who carry such
defects.
Invited papers of the symposium were followed by 21 papers and
demonstrations by members of the Society. The majority of papers presented
new data which were a further but more detailed development of the theme of
the meeting and fell into three categories, cultured cells and organic acid
metabolism, cultured cells and acid hydrolase deficiencies and methodological
developments. These were preceded by invited introductions by D. Gompertz
on organic acid metabolism and by M. F. Niermeijer on methodological
developments. These papers are listed in this monograph and will be published.
Our main aim has been to provide a practical and theoretical background for
the very varied requirements of work on hereditary metabolic disease using
cultured cells. We therefore hope that our attempt to provide a framework for
interdisciplinary communication which has been so well supported by the
authors themselves will be of help in medical practice, research and possibly in
teaching.
We must express the thanks of all involved to Mr J. Milner for his generous
support. We were privileged to have Dr G. Komrower as president, host and
chairman during the meeting. It is also a pleasure to thank the City of Edin
burgh for its warm and generous hospitality which was so much enjoyed by all,
particularly by our many overseas colleagues. This meeting would not have
been possible but for the kindness and efficiency of many members of the staff
of the University of Edinburgh and of its teaching hospitals, especially the
PREFACE ix
Royal Hospital for Sick Children, Edinburgh and the staff of the Biochemistry
Department at Alder Hey Children's Hospital. Our special thanks are also due
to the many people who have helped in the preparation of this book.
Forrester Cockburn
R. Angus Harkness
List of Contributors
A. ADAMS
Department of Paediatric Biochemistry, Royal Hospital for Sick Children, Sciennes Road,
Edinburgh EH9 ILJ
N. R. BELTON
Department of Child Life and Health, University of Edinburgh Medical School, Hatton Place,
Edinburgh EH9 I UW
G. T. N. BESLEY
Department of Pathology, Royal Hospital for Sick Children, Sciennes Road, Edinburgh EH9
ILF
M. E. BLASCOVICS
Children's Hospital, 4650 Sunset Boulevard, Los Angeles, California 90027, USA
K. B. BLAU
Barron Memorial Research Laboratories, Queen Charlotte's Maternity Hospital, Goldhawk
Road, London W6 OXG
N. J. BRANDT
Department of Teratology, Rigshospitalet, Juliane Mariesvej, DK2100 Copenhagen, Denmark
D. M. BROADHEAD
Department of Pathology, Royal Hospital for Sick Children, Sciennes Road, Edinburgh EH9
ILF
EDNA BROWN
Division of Inherited Metabolic Disease, MRC Clinical Research Centre, Northwick Park
Hospital, Watford Road, Harrow, Middlesex
J. BUTTERWORTH
Department of Pathology, Royal Hospital for Sick Children, Sciennes Road, Edinburgh EH9
ILF
NINA CARSON
Nuffield Department of Child Health, Institute of Clinical Science, Grosvenor Road, Belfast
BTl2 6BJ, Northern Ireland
R. A. CHALMERS
Division of Inherited Metabolic Disease, MRC Clinical Research Centre, Northwick Park
Hospital, Watford Road, Harrow, Middlesex
E. CHRISTENSEN
Department of Teratology, Rigshospitalet, Juliane Mariesvej, DK2100 Copenhagen, Denmark
F. COCKBURN
Paediatric Laboratory, Simpson Memorial Maternity Pavilion, Lauriston Place, Edinburgh
EH3 9EF
V. DUBOWITZ
Department of Paediatrics, Royal Postgraduate Medical School, Hammersmith Hospital, Du
Cane Road, London Wl2 OHS
