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Biomedical subjects

J Lehmann

Publications and source records attributed to J Lehmann.

At least 145 records · Page 8Linked to original sources
PubMed1994

Significant inhibition of hybridoma cells by exogenous application of ganglioside GM3, a possible modulator of cell growth in vitro.

Gangliosides of the mouse-rat hybridoma cell line 187.1, which secretes an antibody against kappa-light chain of mouse IgG, were isolated and structurally characterized by biochemical and immunological methods (overlay technique), and fast atom bombardment-mass spectrometry. Exclusively GM3, substituted with C24:1 and C16:0 fatty acid and C18:1 sphingosine, was found in this B cell derived cell line. A GM3(NeuGc) to GM3(NeuAc) ratio (80 to 20), was characteristic for 187.1 cells, and absolute GM3 amounts of about 0.3 mg 10(-9) viable cells were determined. Exogenous application of GM3, which has been isolated from large cell preparations, to 187.1 cells showed growth inhibition in a concentration dependent manner. Using the MTT-assay and the [3H]thymidine incorporation assay, the cells exhibited a strong reduction in metabolic and proliferative activity, respectively, after exposure of cells to GM3. GM3 was applied in concentrations between 3 microM and 30 microM, giving evidence for strong inhibitory effects at 30 microM GM3 and less but significant suppression after application of GM3 concentrations lower than 20 microM. No cellular response was observed at the lowest concentration (3 microM) used in this study. Hybridoma cells as well as other cell types like fibroblasts, muscle cells and endothelial cells, are in general characterized by high expression of the GM3 ganglioside, which is known to act as a modulator of cellular growth in monolayer cultures of adherent cells. Since gangliosides are released to the culture medium by cell lysis, i.e. cell death, and/or by active membrane shedding, the results obtained in this study suggest a growth regulatory role of GM3 in high density hybridoma cell cultures.

Animals↗
PubMed1994

Evaluation of membranes for use in on-line cell separation during mammalian cell perfusion processes.

In this study two microporous hollow fibre membranes were evaluated for their use as cell retention device in continuous perfusion systems. A chemically modified permanent hydrophillic PTFE membrane and a hydrophilized PP membrane were tested. To investigate the filtration characteristic under process conditions each membrane was tested during a long term perfusion cultivation of a hybridoma cell line. In both cultivations the conditions influencing membrane filtration (e.g. transmembrane flux) were kept constant. Filtration behaviour was investigated by monitoring transmembrane pressure and protein permeability. Transmembrane pressure was measured on-line with an autoclavable piezo-resistive pressure sensor. Protein permeability was determined by quantitative evaluation of unreduced, Coomassie stained SDS-PAGE. The membrane fouling process influences the filtration characteristic of both membranes in a different way. After fermentation the PP membrane was blocked by a thick gel layer located in the big outer pores of the asymmetric membrane structure. The hydraulic resistance was higher but the protein permeability was slightly better than of the PTFE membrane. For this reason the PP membrane should be preferred. On the other hand, transmembrane pressure decreases slower when the PTFE membrane is used, which favours this membrane for long term cultivations, especially when low molecular weight proteins (< 30 KD) are produced.

Animals↗
PubMed1994

A direct computer control concept for mammalian cell fermentation processes.

In the last 10 years, new assignments and the special demands of mammalian cells to the culture conditions caused the development of complex small scale fermentation setups. The use of continuous fermentation and cell retention devices requires appropriate process control systems. An arrangement for control and data-acquisition of complex laboratory-scale bioreactors is presented. The fundamental idea was the usage of a standard personal computer, which is connected to pumps, valves and sensors via ADA-transformation. The possibility of free programming allowed the development of user-oriented software, especially designed for the far-reaching requirements of a university laboratory in the field of animal cell culture. Control of aeration, pumps, data-acquisition and data-storage are combined within one program, which allows the automation of standard operations like measurement of kLa- or OTR-values. Pump control algorithms for all common fermentation strategies (batch, fed batch, chemostat, perfusion) are included and can be selected any time during cultivation. Oxygen partial pressure and pH are controlled via direct digital control (ddc), providing simple adaption of control parameters and set points to current fermentation conditions.

Animals↗
PubMed1994

Design of a bubble-swarm bioreactor for animal cell culture.

A stationary bubble-swarm has been used to aerate a mammalian cell culture bioreactor with an extremely low gas flow rate. Prolonging the residence time of the gas bubbles within the medium improved the efficiency of the gas transfer into the liquid phase and suppressed foam formation. An appropriate field of speed gradients prevented the bubbles from rising to the surface. This aeration method achieves an almost 90% transfer of oxygen supplied by the bubbles. Consequently, it is able to supply cells with oxygen even at high cell densities, while sparging with a gas flow of only 0.22 x 10(-3) -1.45 x 10(-3) vvm (30-200 ml/h). The reactor design, the oxygen transfer rates and the high efficiency of the system are presented. Two repeated batch cultures of a rat-mouse hybridoma cell line are compared with a surface-aerated spinner culture. The used cell culture medium was serum-free, either with or without BSA and did not contain surfactants or other cell protecting agents. One batch is discussed in detail for oxygen supply, amino acid consumption and specific antibody production.

Ammonia↗
PubMed1994

The Super-Spinner: a low cost animal cell culture bioreactor for the CO2 incubator.

The production of small quantities of monoclonal antibodies and recombinant proteins was carried out using a new low cost production system, the Super Spinner. Into a 1 1 standard Duran flask a membrane stirrer equipped with a polypropylene hollow fiber membrane was installed to improve the oxygen supply by bubble-free aeration. The aeration was facilitated by using the CO2 conditioned incubator gas, which was pumped through the membrane stirrer via a small membrane pump. The maximal oxygen transfer rate (OTRmax) of the Super Spinner was detected. For this purpose one spinner flask was equipped with an oxygen electrode. The OTRmax was measured by the dynamic method. The ratio of membrane length to culture volume was adapted corresponding to the oxygen uptake rate of the cells according to the desired cell density. A balanced nutrient supply resulted in an optimal formation and yield of products.

Animals↗
PubMed1994

Expression of gangliosides GM3 (NeuAc) and GM3 (NeuGc) in myelomas and hybridomas of mouse, rat, and human origin.

In this study gangliosides from various myelomas and hybridomas of mouse, rat, and human origin were characterized by thin-layer and high-performance liquid chromatography, immunological methods (overlay technique) and fast atom bombardment mass spectrometry. Exclusively GM3 substituted with C24:1- and C16:0-fatty acid, was found in all B cell-derived cell lines. C18 sphingosine was the single long chain base in each GM3 ceramide portion. The mouse myeloma (NS-1) and all hybridomas, obtained by fusion of mouse, rat, or human B lymphocytes with murine myelomas, showed high GM3 (NeuGc) content (> 75%) and low GM3 (NeuAc) expression. Absolute amounts of GM3 ranged from 0.2 up to 0.8 mg x 10(-9) cells. Normally, human cells do not express NeuGc, and an Epstein-Barr virus-transformed human B lymphocyte line analyzed in this study retained this sialylation status, expressing exclusively GM3 (NeuAc) (100%). The fusion of human B lymphocytes with mouse myelomas led to high GM3 (NeuGc) expression (average about 85%) in all mouse/human heterohybridomas examined. Our results indicate the chromosomal gene "transfer" and/or the activation of enzymes involved in NeuGc-biosynthesis due to the somatic cell fusion process, which might explain the mouse dominance in the manifestation of the NeuGc-phenotype in hybridomas of human origin.

Animals↗
PubMed1993

Synthesis and evaluation of diastereoisomeric alkylating pseudo-disaccharides as potential affinity reagents for trehalase.

Reaction of (+/-)-(3/4,5,6)-4-bromo-5,6-epoxy-3-hydroxycyclohexene with 2,3,4,6-tetra-O-acetyl-1-thio-alpha-D-glucopyranose, followed by treatment of the resulting isolated diastereoisomeric 4-bromo-3,5-dihydroxycyclohexene 1-thioglycoside derivatives with base under phase-transfer conditions, gave (R)- and (S)-(3,4,6/5)-3,4-epoxy-6-S-(1-thio-alpha-D-glucopyranosyl)-5- hydroxycyclohexene. None of them was substrate or inhibitor for cockchafer trehalase.

Alkylation↗
PubMed1993

New amino sugar analogues are incorporated at different rates into glycoproteins of mouse organs.

Different radiolabelled N-acyl-derivatives of D-glucosamine were synthesized using D-glucosamine and the respective carbonic acid anhydride. Metabolism of these sugar analogues could be shown in vitro as well as in vivo. After the intraperitoneal administration of these radiolabelled N-acyl-D-glucosamines to mice, their rate of incorporation into glycoproteins of different organs was found to increase markedly with the length of the N-acyl side chain. Highest incorporation was measured in the whole intestine using N-pentanoyl-D-glucosamine as label.

Acetylglucosamine↗
PubMed1993

Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant.

The task of purifying monoclonal antibodies (MAbs) and human recombinant antithrombin III (rATIII) from cell culture supernatant was carried out using two different approaches, both based on the use of membraneous matrices. The first approach employed a strongly acidic and a strongly basic membrane ion exchanger, which were evaluated for their ability to purify monoclonal antibodies and the human active recombinant antithrombin III from cell culture supernatant. Within minutes gram amounts of product could be purified in a high-flux system, specially developed for this purpose, achieving purities of 80% for MAbs and 75% for rATIII, respectively. The capacity of the acidic membrane ion exchanger for MAbs was found to be 1 mg/cm2 with recoveries up to 96% and that of the basic membrane ion exchanger for rATIII was 0.15 mg/cm2 with recoveries up to 91%. The second approach consisted of using heparin, a mucopolysaccharide with a strong affinity towards ATIII, coupled to amine-modified or epoxy-activated membranes by reductive amination, for the purification of rATIII. The ATIII binding capacities of the membranes were found to be 91 micrograms/cm2 for the amine-modified and 39 micrograms/cm2 for the epoxy-activated membrane, achieving purities of 75%. The coupling proved to be fairly stable over a period of 5 months and the membranes remained operable even after steam sterilization and treatment with sodium dodecyl sulphate. Final purification in both instances was carried out by gel filtration.

Animals↗
PubMed1993

[Lactones. 24. "Gamma-lactonized" neuroleptics--synthesis, stereochemistry and affinity at the D2 receptor].

Addition of piperidines 15a-d to methylene lactones 8,14a gives the "gamma-lactonized" diphenylbutylpiperidine-type neuroleptics 3a,b,d,4a-d. 8 can be produced by cyclization or--as well as 14--from the corresponding alpha-carboxylactone. 1H- and 13C-nmr data give evidence for the equatorial position of the aminomethyl group in all derivatives of 3 and 4 and bisequatorial-trans-configuration in 4a-d. 3H-spiroperidole displacement at the D2 receptor in rat brain by 3,4 and the "gamma-lactonized" butyrophenone neuroleptics 1,2a-c was investigated. Some compounds show moderate up to high (3b) activity.

Animals↗
PubMed1993

[Indoles. 9. 4-Arylated tetrahydro-beta-carbolines--synthesis and first pharmacologic data].

Two different routes lead to the 4-arylated tetrahydro-beta-carbolines 5, 11, 12. One includes Pictet-Spengler cyclization of tryptamines 9, the other follows our "lactone-route", starting with 1 via 2-4. In a first pharmacological screening some target compounds show significant affinity at the 5-HT2-receptor but no or only low affinity to other binding sites.

Animals↗
PubMed1993

[Morphologic causes and consequences of fracture of the distal radius].

Age and stress related changes in the distal epiphysis of the radius are viewed as cause and effect of losses in stability due to pathological stress. 15 in 97 distal radii show typical osseous alterations which can be evidenced after healing of a radial fracture by means of determining the subchondral cortex density. A dorsally directed inclination of the distal radius epiphysis following a healed fracture often be observed. This inclination leads to a redirecting of the resultant force vector which in turn results in a thickening of the subchondral corticalis directed towards the palmar socket margin. The densitometric evaluation based on X-ray depictions enables the morphological characterisation of the restructuring of corticalis and spongiosa in long bones as cause and effect of a fracture.

Aged↗