New Developments and New Applications in Animal Cell Technology

by Merten, Otto-Wilhelm; Perrin, Pierre; Griffiths, Bryan

• ISBN13: 9780792350163
• ISBN10: 0792350162
• Format: Hardcover
• Copyright: 1998-11-01
• Publisher: Kluwer Academic Pub

Summary

Animal cell technology is becoming an increasingly important part of biotechnology and many products are now used in human health care and for veterinary applications. However, there are many times more products actually in the developmental pipelines of the biotechnology industry, including various phases of clinical trials. The Proceedings of the 15th Meeting of the European Society for Animal Cell Technology (Tours, France, September 1997) presents the actual current state as well as New Developments and Applications in Animal Cell Technology for the benefit of society. These Proceedings represent both the current state and applications of animal cell technology and the way the technology is expanding into new areas to give a unique insight into new products and applications for human and animal health care.

Table of Contents

Introduction	xxiii
HYCLONE Lecture
Genetic manipulation of the mammalian genome (Abstract)	1	(4)
R. Jaenisch
ESACT Lecture
Engineering glycosylation in animal cells	5	(24)
J.E. Bailey
E. Prati
J. Jean-Mairet
A. Sburlati
P. Umana
Sessions on:
Biosynthesis and post-translational modifications of recombinant proteins, and Cell physiology and metabolic engineering of animal cells	25	(4)
Gene expression:
Enhanced recombinant protein expression in insect cells in the presence of dimethylsulfoxide (DMSO)	29	(6)
L. Ramos
J.F. Kane
A.A. Murnane
Studies of the hepatitis C virus recombinant protein production and its effects on Sf-9 insect cells metabolism	35	(4)
C.M. Charon
Characterization of phosphorylation mutants of the human multidrug transporter (MDR1) expressed in the baculovirus-Sf9 insect cell system	39	(4)
K. Szabo
E. Bakos
E. Welker
A. Varadi
H.R. Goodfellow
C.F. Higgins
B. Sarkadi
Glutamine synthetase transfected cells may avoid selection by releasing glutamine	43	(8)
P. Bird
E. Bolam
L. Castell
O. Obeid
N. Darton
G. Hale
Study of different cell culture conditions for the production of a « reshaped » Mab in NSO cells	51	(4)
A.J. Castillo
A. Fernandez
T. Boggiano
P. Pugeaud
I.W. Marison
Increasing monoclonal antibody productivity by semicontinuous substitution of production medium for growth medium	55	(4)
F. Franek
K. Sramkova
Expression of recombinant human insulin in chinese hamster ovary cells is complicated by intracellular insulin-degrading enzymes	59	(10)
S.C.O. Pak
S.M.N. Hunt
M.J. Sleigh
P.P. Gray
High yield expression of recombinant plasma factors: use of recombinant endoprotease derivatives in vivo and in vitro	69	(8)
U. Schlokat
A. Preininger
M. Himmelspach
G. Mohr
B. Fischer
F. Dorner
The role of cell cycle in determining levels of gene expression in CHO cells	77	(4)
D.R. Lloyd
V. Leelavatcharamas
D.C. Edwards
A.N. Emery
M. Al-Rubeai
Heterogeneity within DHFR-mediated gene amplified population of CHO cells producing chimeric antibodies	81	(4)
N.S. Kim
S.J. Kim
G.M. Lee
Immunofluorescence detection of recombinant monoclonal antibody as a tool in the characterization of chinese hamster ovary cell lines	85	(8)
C.A.S. Kinney
R.E. Trala
L.C. Hendricks
T.D. Hill
Electrolyzed reduced water which can scavenge active oxygen species supresses cell growth and regulates gene expression of animal cells	93	(4)
S. Shirahata
S. Kabayama
K. Kusumoto
M. Gotoh
K. Teruya
K. Otsubo
T. S. Morisawa
H. Hayashi
K. Katakura
Differential gene expression of cytochrome P450 in immortalised hepatocyte cell lines	97	(4)
H.T. Kelly
K. Anderson
E. Hill
H. Grant
C. MacDonald
Post-transcriptional control of recombinant gene expression: mRNA retargeting and regulation of expression	101	(4)
K.A. Partridge
A. Johannessen
A. Tauler
I.F. Pryme
J.E. Hesketh
Transient transfection in mammalian cells. A basic study for an efficient and cost-effective scale-up process	105	(8)
E.-J. Schlaeger
L.Y. Legendre
A. Trzeciak
E.A. Kitas
K. Christensen
U. Deuschle
A. Supersaxo
Large scale transient gene expression in mammalian cells	113	(4)
E.-J. Schlaeger
K. Christensen
G. Schmid
N. Schaub
B. Wipf
A. Weiss
Synergistic enhancement of transient expression by dioleoyl-melittin (DOM) and polyethylenimine (PEI) in mammalian cells in suspension culture	117	(4)
E.-J. Schlaeger
J.Y. Legendre
A. Trzeciak
E.A. Kitas
K. Christensen
U. Deuschle
A. Supersaxo
Transient expression of a soluble and secreted form of heterodimeric T-cell receptor in HEK-293	121	(4)
M. Jordan
C.L. Blanchard
I. Bernasconi
I. Luescher
F.M. Wurm
Measurable parameters of cells and precipitate predict transfectability with calcium phosphate	125	(4)
M. Jordan
F. Wurm
Glycosylation:	129	(37)
Influence of Na-butyrate on the production and sialylation of human interferon-g by a2,6-sialyltransferase engineered CHO-cells	131	(4)
D. Lamotte
L. Monaco
N. Jenkins
A. Marc
Elevated pCO2 inhibits the polysialylation of the neural cell adhesion molecule in CHO MT2-1-8 cell cultures	135	(6)
J.A. Zanghi
T.P. Mendoza
R.H. Knop
W.M. Miller
Influence of cultivation conditions on glycosylation pattern - a fed-batch and continuous culture study	141	(8)
N.A. Schill
M.Z. Rosenberg
R.L. Dabora
Effects of different production systems on glycosylation pattern of murine monoclonal IgA	149	(4)
F. Schweikart
E. Lullau
R. Jones
G.J. Hughes
Identification of altered glycosylation as the major difference between intracellulary accumulated and secreted b-trace protein produced in baculovirus-infected insect cells	153	(4)
G. Rodriguez
H.S. Conradt
V. Jager
How ammonium dominates the metabolism of in vitro cultivated mammalian cells	157	(6)
A. Cayli
M. Wirth
R. Wagner
Study of human recombinant GM-SCF produced in different host systems using monoclonal antibodies	163	(3)
M. Etcheverrigaray
M. Oggero
M. Bollati
R. Kratje
Metabolic engineering:	166	(29)
Modification of hybridoma cells metabolism	167	(8)
J.J. Cairo
C. Paredes
F. Godia
E. Prats
F. Azorin
Ll. Cornudella
Cloning and expression of a cytosolic sialidase from CHO cells in a glutathione S-transferase (GST)-encoding expression vector	175	(6)
M. Burg
J. Muthing
Construction of a novel CHO cell line coexpressing human glycosyltransferases and fusion PSGL-1 - immunoglobulin G	181	(4)
B. Vonach
B. Hess
C. Leist
Production of defined glycosylation variants of secreted human of secreted human glycoprotein therapeutics by coexpression with human recombinant glycosyltransferases in BHK-21 cells	185	(6)
P. Schlenke
E. Grabenhorst
J. Costa
M. Nimtz
H.S. Conradt
Antisense RNA for the elimination of NeuGc residues from recombinant glycoproteins	191	(4)
A. Gregoire
A. Visvikis
A. Marc
J.-L. Goergen
Cell proliferation, proliferation control, apoptosis:	195	(70)
Intracellular fatty acid composition affects cell yield, energy metabolism and cell damage in agitated cultures	197	(8)
M. Butler
N. Huzel
N. Barnabe
L. Bajno
T. Gray
Lipid requirements of a recombinant chinese hamster ovary cell line (CHO)	205	(4)
J.T. McGrew
C.L. Richards
P. Smidt
B. Dell
V. Price
Sustained expression in proliferation controled BHK-21 cells	209	(6)
P.P. Muller
S. Kirchhoff
H. Hauser
Cell growth inhibition by the IRF-1 system	215	(4)
A.V. Carvalhal
J.L. Moreira
P. Muller
H. Hauser
M.J.T. Carrondo
Correlation between BHK cell specific productivity and metabolism	219	(4)
H.J. Cruz
J.L. Moreira
E.M. Dias
C.M. Peixoto
A.S. Ferreira
M.J.T. Carrondo
Effect of growth arrest in BHK metabolism: on line monitoring by 13C and 31P NMR spectroscopy	223	(4)
P.M. Alves
A.V. Carvalhal
J.L. Moreira
H. Santos
M.J.T. Carrondo
Cell survival in CHO cell cultures grown in a defined protein free medium	227	(4)
N. Chatzisavido
C. Fenge
L. Haggstrom
Enhanced apoptosis in insect cells cultivated in simulated microgravity	231	(4)
N. Cowger
K. O'Connor
Modulation of apoptosis by bcl-2 expression following amino acid deprivation and in high cell density perfusion cultures	235	(8)
R.P. Singh
D. Fassnacht
A. Perani
N.H. Simpson
C. Goldenzon
R. Portner
M. Al-Rubeai
Effect of bcl-2 expression on hybridoma cell growth during stressful conditions	243	(4)
D. Fassnacht
S. Rossing
F. Franek
M. Al-Rubeai
R. Portner
Inhibition of c-jun expression in F-MEL cells causes cell cycle arrest and prevention of apoptosis	247	(8)
Y.H. Kim
T. Iida
E.V. Prochownik
E. Suzuki
Hippocampal cells in culture as a model to study neuronal apoptosis	255	(4)
I. Figiel
J. Jaworski
L. Kaczmarek
Development of carboxy SNARF-1-AM and annexin V assays for the determination of apoptosis in heterogeneous cultures	259	(6)
A. Ishaque
M. Al-Rubeai
Session on:
Integrated bioprocessing in animal cell technology	263	(2)
Physical environment, cells and products:	265	(42)
Diauxic cell behavior enables detoxification of CHO cell culture medium during fed batch cultivation	267	(6)
H. Lubben
G. Kretzmer
Effects of polysaccharide derived from tea on growth of human cell lines in serum free culture	273	(4)
H. Kawahara
M. Maeda-Yamamoto
K. Osada
K. Tsuji
Metabolic demands of BTI-TN-5B1-4 (High Five™) insect cells during growth and after infection with baculovirus	277	(4)
E. Chico
V. Jager
Effects of ammonium and lactate during continuous hybridoma fermentations in a fluidized-bed bioreactor	281	(4)
H. Heine
M. Spies
M. Biselli
C. Wandrey
Serum concentration and pH affect the CD13 receptor content of HL60 cells cultured in stirred bioreactors	285	(8)
C.L. McDowell
E.T. Papoutsakis
Four regulation-friendly serum-free media for mammalian and insect cells	293	(10)
T.W. Irish
S.E. Lenk
L.A. Bugner
K.J. Etchberger
Proteolytic activities in the baculovirus-insect cell expression system	303	(4)
G. Schmid
A. Bischoff
Monitoring and control:	307	(60)
Development of proteinase assays for improved CHO cell cultures	309	(8)
C. Tans
C. vander Maelen
S. Wattiaux-de Coninck
M.-M. Gonze
L. Fabry
Digital image analysis: quantitative evaluation of colored microscopic images of animal cells	317	(4)
K. Falkner
E.D. Gilles
The viable cell monitor: a dielectric spectroscope for growth and metabolic studies of animal cells on macroporous beads	321	(8)
Y. Guan
R.B. Kemp
Measurements of changes in cell size distribution to monitor baculovirus infection of insect cells	329	(4)
E. Chico
V. Jager
Dead cell estimation - a comparison of different methods	333	(4)
A. Falkenhain
Th. Lorenz
U. Behrendt
J. Lehmann
Fed-batch culture development based on biomass monitoring	337	(6)
E. de Buyl
A. Maxwell
L. Fabry
On-line monitoring of protein and substrate/product concentration in mammalian cell cultivation	343	(4)
H. Lubben
J. Hagedorn
T. Scheper
Process control and on-line feeding strategies for fed-batch and dialysis cultures of hybridoma cells	347	(4)
J.O. Schwabe
R. Portner
Metabolic network analysis of a hybridoma cell line using mass balances and C14-labelled glucose and glutamine	351	(4)
P.-A. Ruffieux
I.W. Marison
U. von Stockar
Dynamic medium optimization by on-line heat flux measurement and a stoichiometric model in mammalian cell culture	355	(4)
Y. Guan
R.B. Kemp
Modeling of glycoprotein production by chinese hamster ovary cells for process monitoring and control	359	(4)
J. Stelling
R.K. Biener
J. Haas
G. Oswald
D. Schuller
W. Noe
E.D. Gilles
The temperature effect in mammalian cell culture. An Arrhenius interpretation	363	(4)
G. Kretzmer
T. Buch
K. Konstantinov
D. Naveh
Integrated processe and scale-up:	367	(100)
Dielectrophoretic forces can be exploited to increase the efficiency of animal cell perfusion cultures	369	(8)
N. Kalogerakis
A. Docoslis
L.A. Behie
Use of a new microcarrier with two-dimensional geometry for the culture of anchorage-dependent cells in sonoperfused, continuously stirred tank reactors	377	(4)
C. Gatot
F. Trampler
M.-P. Wanderpepen
J. Harfield
A. Oudshoorn
A. Johansson
V. Nielsen
A.O.A. Miller
Evaluation of porous microcarriers in fluidized bed reactor for protein production by HEK 293 cells	381	(4)
M.A. Valle
J. Kaufman
W.E. Bentley
J. Shiloach
Fluidized bed technology: influence of fluidization velocity on nutrient consumption and product expression	385	(4)
G. Bluml
M. Reiter
Th. Gaida
N. Zach
A. Assadian
C. Schmatz
H. Katinger
High density and scale-up cultivation of recombinant CHO cell line and hybridomas with porous microcarrier cytopore	389	(6)
C. Xiao
Z. Huang
W. Li
X. Hu
W. Qu
L. Gao
G. Liu
Cell-settler perfusion system for the production and glycosylation of human interferon-g by clumped cells	395	(4)
D. Lamotte
J. Straczek
A. Marc
Disposable bioreactor for cell culture using wave-induced agitation	399	(10)
V. Singh
Use of miniPERM system for an efficient production of mouse monoclonal antibodies	409	(8)
M.L. Nolli
R. Rossi
A. Soffientini
D. Zanette
C. Quarta
Production of anti-EGF receptor hMAb in hollow fiber bioreactors for in vivo diagnosis	417	(4)
M.A. Arias
A. Valdes
D. Curbelo
O.M. Morejon
I. Caballero
J. Villan
J.A. Gomez
A. Fernandez
J. Rodriguez
A. Morales
T. Boggiano
A. Castillo
L. Bouzo
C. Hermida
G. Garcia
P. Viton
N. Perez
T. Rodriguez
Optimisation of the production of VLP's in 2 Lt stirred bioreactors	421	(8)
P.E. Cruz
A. Cunha
C.C. Peixoto
J. Clemente
J.L. Moreira
M.J.T. Carrondo
Direct capture of monoclonal antibodies with a new high capacity streamline SP XL ion exchanger	429	(4)
C. Priesner
N. Ameskamp
D. Lutkemeyer
J. Lehmann
The effect of different purification schemes on the activity of a monoclonal antibody	433	(4)
D.J. Black
J.P. Barford
C. Harbour
A. Fletcher
Interfacing of protein product recovery with an insect cell baculovirus production system	437	(4)
I.A. Carmichael
M. Al-Rubeai
A. Lyddiatt
Preparation of plasmid DNA using displacement chromatography	441	(4)
R. Freitag
S. Vogt
Novel methods for large-scale preparation of nutrient media and buffered salt solutions	445	(4)
D.W. Jayme
Large scale application of the Semliki Forest Virus system: 5-HT3 receptor production	449	(8)
H.D. Blasey
B. Brethon
R. Hovius
K. Lundstrom
L. Rey
H. Vogel
A.-P. Tairi
A.R. Bernard
From applied research to industrial application: The success story of monitoring intracellular ribonucleotide pools	457	(6)
S.I. Grammatikos
K. Tobien
W. Noe
R.G. Werner
New technology of production recombinant human erythropoietin for peroral administration	463	(4)
T.D. Kolokoltsova
N.E. Kostina
E.A. Nechaeva
N.D. Yurchenko
O.V. Shumakova
A.B. Rizikov
N.A. Varaksin
T.G. Ryabicheva
General safety aspects:	467	(298)
Significance of multiple testing on murine leukemia virus of mouse hybridomas	469	(4)
E.J.M. Al
T. Jorritsma
A. Blok
H.M.G. Sillekens
P.C. van Mourik
Murine retrovirus detection using Mus dunni cells S+L- cells	473	(8)
P. Seechurn
B. Mortimer
P. Newton
C. Martin
The removal of model viruses during the purification of human albumin using chromatographic procedures	481	(4)
R. Cameron
C. Harbour
Y. Cossart
J.P. Barford
Inactivation of hepatitis A virus during the production of Nordimmun	485	(8)
D.W. Birch
P. Kaersgaard
C. Martin
Session on:
Viral vector production for gene therapy	491	(2)
Adeno-associated viral vectors: Principles and in vivo use	493	(10)
O. Danos
Novel retroviral packaging cell lines: Improved vector production for efficient hematopoietic progenitor cell transduction	503	(6)
S. Forestell
R. Rigg
J. Dando
J. Chen
B. Joyce
R. Tushinski
C. Reading
E. Bohnlein
Scale-up and otpimisation issues in gene therapy. Viral vector production	509	(6)
J.E. Boyd
L. Borland
C.E. Fisher
Comparison of manufacturing techniques for adenovirus production	515	(8)
J.M. Ostrove
P. Iyer
J. Marshall
D. Vacante
Safety considerations in the development of new retroviral and adenoviral vectors for gene therapy	523	(8)
D. Morgan
I. Forgie
J. Ostrove
M.H. Wisher
Gene therapy for neuromuscular disorders: Macrophages as shuttles for widespread targeting	531	(10)
E. Parrish
E. Peltekian
L. Garcia
Gene transfer into dog myoblasts	541	(4)
S. Braun
C. Thioudellet
C. Escriou
M.-C. Claudepierre
F. Langle-Rouault
E. Jacobs
R. Bischoff
D. Elmlinger
H. Homann
Y. Poitevin
M. Lusky
M. Mehtali
F. Perraud
A. Pavirani
Biodistribution analysis of a gene therapy vector using the polymerase chain reaction (PCR) technique	545	(6)
J. Proffitt
M. Leibbrandt
K. Jarvis
C. Martin
Session on:
Developments for immunologicals and vaccines	549	(2)
The production of influenza virus by immobilised MDCK cells	551	(4)
D. Looby
J. Tree
A. Talukder
K. Hayes
H. House
G. Stacey
Serum-free grown MDCK cells: An alternative for influenza vaccine virus production	555	(6)
N. Kessler
G. Thomas
L. Gerentes
M. Aymard
Evaluation of the new medium (MDSS2N), free of serum and animal proteins, for the production of biologicals	561	(8)
H. Kallel
P. Perrin
O.-W. Merten
Production of high titre disabled infectious single cycle (DISC) HSV-2 from a microcarrier culture	569	(4)
T.A. Zecchini
R.J. Smith
New form of the live measles vaccine for oral administration	573	(4)
E.A. Nechaeva
E.A. Kashentseva
A.P. Agafonov
N.A. Varaksin
T.G. Ryabicheva
A.P. Konstantinov
V.N. Bondarenko
T.D. Kolokoltsova
I.V. Kits
T. Yu. Sen'Kina
N.V. Zhilina
Study of Leningrad-16 vaccine strain of measles virus reproduction in cell cultures perspective for biotechnology	577	(4)
E.A. Nechaeva
T.N. Getmanova
T.Y. Sen'Kina
N.D. Yurchenko
Recent advances with new vaccine adjuvants from preclinical to clinical development (Abstract)	581	(2)
T. Voss
Vaccination with recombinant suicidal DNA/RNA	583	(10)
P. Berglund
M. Fleeton
C. Smerdou
I. Tubulekas
B.J. Sheahan
G.J. Atkins
P. Liljestrom
Growth of goat endothelial cells for the production of a veterinary vaccine	593	(4)
P.M. Miranda
J.L. Moreira
M.J.T. Carrondo
Expression in insect cells of the major parasite antigen associated with human resistance to Schistosomiasis	597	(4)
L. Argiro
C. Doerig
S. Liabeuf
A. Bourgois
J.L. Romette
Modulation of CD4 expression on helper T lymphocytes and U937 cells by ganglioside GM3 and its derivatives	601	(6)
D. Heitmann
P. Budde
J. Frey
J. Lehmann
J. Muthing
Human serum in leukocyte cultures producing human interferon alpha	607	(6)
P. Mattana
L. Scapol
S. Silvestri
G.C. Viscomi
The new type of immunomodulator	613	(4)
M.V. Mezentseva
V.A. Mozgovoi
L. Yu. Mozgovaya
R. Ya. Podchernyaeva
In vitro immunization of human peripheral blood lymphocytes with cholera toxin B subunit	617	(10)
A. Ichikawa
Y. Katakura
T. Kawahara
S. Hashizume
S. Shirahata
Session on:
New technologies for health care products	625	(2)
Analysis of cell growth in a fixed bed bioreactor using magnetic resonance spectroscopy and imaging	627	(8)
P.E. Thelwall
M.L. Anthony
D. Fassnacht
R. Portner
K.M. Brindle
Expansion of human hematopoietic progenitor cells in a fixed bed bioreactor (Abstract)	635	(2)
P. Meissner
P. Werner
B. Schroder
C. Herfurth
C. Wandrey
M. Biselli
A novel assay to determine and quantify the regulation of cell motility and migration demonstrated on hematopoietic cells	637	(8)
D. Mobest
S. Ries
R. Mertelsmann
R. Henschler
Immortalization of differentiated hepatocytes	645	(12)
G.S. Jennings
M. Strauss
Fixed-bed reactors for animal cell cultivation: An approach to artificial organs	657	(4)
R. Portner
S. Rossing
J. Stange
D. Fassnacht
High density perfusion culture of primary rat hepatocytes for potential use as a bioartificial liver device	661	(4)
K. Bratch
A.J. Strain
M. Al-Rubeai
Metabolic competence and hormonal regulation of primary porcine hepatocytes in a 3-D sandwich configuration (Abstract)	665	(4)
A. Bader
D. Rocker
A. Acigkoz
S. Schwintek
J. von Schweder
M. Maringka
V. Armstrong
R. Wagner
G. Steinhoff
A. Haverich
Novel mini-bioreactors for islet cell culture	669	(4)
A. Handa-Corrigan
I.C. Green
J. Mabley
S. Hayavi
G.N. Kass
R.H. Hinton
L.M. Morgan
J. Wright
Cultivation of skin cells suitable for recovery of burn wounds	673	(4)
T.D. Kolokoltsova
N.D. Yurchenko
N.G. Kolosov
O.V. Shumakova
E.A. Nechaeva
Tissue therapy for treatment of primary myodystrophies	677	(4)
T.B. Krokhina
G.B. Raevskaja
S.S. Shishkin
Possibility of application of thyroid organ culture for the treatment of persistent hypothyroidism	681	(4)
I.P. Pasteur
N.D. Tronko
E.N. Gorban
V.I. Kravchenko
ESACT Lecture
Human cells as therapeutic agents (Abstract)	685	(4)
H. Green
Session on:
Use of animal cells for in vitro testing	687	(2)
Embryonic stem cell differentiation models: Cardio-vascular, myogenic and neurogenic development in vitro	689	(16)
A.M. Wobus
K. Guan
J. Rohwedel
C. Strubing
M. Drab
Responses of human lung epithelial cells (A549) to pathogenic infection by Mycoplasma pneumoniae	705	(8)
J. Goodman
K. Morley
P. Packer
T. Battle
Polar lipid profiling of Mycoplasma pneumoniae-infected human lung epithelial cells	713	(4)
J. Goodman
R. Wait
T. Battle
Comparison of three dimensional (3-D) rat hepatocyte cultures in simulated microgravity conditions	717	(4)
T. Maguire
H.J. Moulsdale
G. Stacey
T. Battle
High density culture of the human hepatoma cell line HepG2: Long-term culture for in vitro toxicology	721	(4)
A. Handa-Corrigan
R.M. Traynor
I. Adamopoulos
J. Salway
Application of primary cultures of rat fetal neurons to the study of neurotrophic action of peptides	725	(4)
O.V. Dolotov
I.A. Grivennikov
Development and validation of an image analysis system for single cell characterization in cell monolayers	729	(4)
D. Kaiser
M.A. Freyberg
G. von Wichert
P. Marenbach
H. Tolle
P. Friedl
Development of an optically accessible perfusion chamber for in situ assays and for long-term cultivation of mammalian cells	733	(4)
M.A. Freyberg
P. Friedl
Analysis of mitogenic activity of proteins after separation by gel electrophoresis	737	(6)
O. Hohenwarter
G. Marzban
E. Jisa
H. Katinger
Workshop on:
The use of animal cells versus the use of transgenic animals for the production of recombinant proteins	741	(2)
Secretion of fusion proteins into milk by transgenic mouse mammary epithelium (Abstract)	743	(2)
D. Pollok
L. Chen
H. Liem
B. Wilburn
J. Williams
M. Harrington
Y. Echelard
H. Meade
The production of proteins in the milk of transgenic livestock: A comparison of microinjection and nuclear transfer	745	(6)
I. Garner
Creation of mice expressing human antibody by introduction of a human chromosome	751	(6)
H. Yoshida
K. Tomizuka
H. Uejima
H. Kugoh
K. Satoh
A. Ohguma
M. Hayasaka
K. Hanaoka
M. Oshimura
I. Ishida
Transgenic technology - A challenge for mammalian cell culture production systems	757	(8)
R.G. Werner
General Workshop-Discussion	765	(8)
List of trade-fair-sponsors	773	(2)
Index	775