Induced differentiation of a neuroblastoma

Schubert, David; Humphreys, Susie; Vitry, F. De; Jacob, François

doi:10.1016/0012-1606(71)90004-2

Cited by 226 publications

(64 citation statements)

References 5 publications

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“…Previous investigations have suggested that under certain conditions morphological differentiation need not be associated with electrical or biochemical differentiation. An example of the former case is that of serum removal, which although inducing the formation of extensive neurites in a number of different neuroblastoma clones (24,25), results in only a minor enhancement in active electrical properties (3). A corresponding dissociation of neurospecific biochemical function from changes in morphology occurs when clone NIE-115 cells are treated with sodium butyrate (7,26), which causes an induction of TH while cells retain a round morphology.…”

Section: Discussionmentioning

confidence: 99%

Maturation of neuroblastoma cells in the presence of dimethylsulfoxide.

Kimhi¹,

Palfrey²,

Spector³

et al. 1976

Proc. Natl. Acad. Sci. U.S.A.

249

144

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Addition of dimethylsulfoxide at concentrations of 1% and 2% (vol/vol) to cells of mouse neuroblastoma clone NIE-115 in the confluent phase of growth resulted in the production of morphologicalfir differentiated cultures with extensive process formation. Cells maintained in 2% dimethylsulfoxide remained in a stable nondividing condition for periods of up to 4 weeks. A high degree of electrical excitability was found in these cells, but there was no clear correlation of this property with the level of induction of either acetylcholinesterase (acetylcholine hydrolase; EC 3.1.1.7) or tyrosine hydroxylase [L-tyrosine, tetrahydropteridine:oxygen oxidoreductase (3-hydroxylating); EC 1.14.16.2]. In addition, intracellular levels of cyclic 3':5'-AMP were not elevated in fully morphologically and electrically differentiated cells. While cell division was markedly inhibited by 2% or higher concentrations of dimethylsulfoxide, at 1% growth continued at a somewhat slowed rate and such cultures exhibited enhanced process formation and electrical activity for a relatively short period. High concentrations (3% or 4%) of dimethylsulfoxide totally suppressed process formation and did not result in increased excitability, but cells maintained high resting potentials. The results suggest that the development of the excitable membrane in neuroblastoma cells maybe expressed independently of neurospecific enzyme induction, and does not require a sustained elevation of cyclic 3'i5'-AMP levels.Nerve cells are characterized by their unique morphological appearance, the possession of an excitable membrane, and a specialized biochemical machinery. In order to study the expression of specific neuronal properties during the maturation process it is necessary to obtain sufficient quantities of a relatively homogeneous differentiating population. Cloned cell lines isolated from the C-1300 mouse neuroblastoma may serve as a useful system for exploring certain aspects of nerve cell differentiation. The transition of a culture of neuroblastoma cells from the actively dividing state to the confluent one is characterized by the synthesis of various enzymes involved in neurotransmitter metabolism (1), an enhancement of electrical excitability (2, 22), and some degree of process formation. To achieve a further expression of these properties, cells have been treated with various agents such as aminopterin (3, 4) or dibutyryl cAMP (5-7).In the present study, we show that in the presence of dimethylsulfoxide (Me2SO) neuroblastoma cells will extend neurites and develop a highly excitable membrane. It appears that this method offers certain advantages over those most commonly used, especially in that cells appear to reach a higher level of electrical differentiation and can be maintained in this state for extended periods. In experiments where logarithmically growing cells were used, cultures were trypsinized before attaining confluency and replated in 60 mm dishes at a density of 1 X 105 cells per dish. Two days later, the medium was replaced w...

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Section: Discussionmentioning

confidence: 99%

Maturation of neuroblastoma cells in the presence of dimethylsulfoxide.

Kimhi¹,

Palfrey²,

Spector³

et al. 1976

Proc. Natl. Acad. Sci. U.S.A.

249

144

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“…Furthermore, B104 cells, like C1300 neuroblastoma cells (11,12), respond to removal of serum by rapidly extending neurites, a phenomenon that has been correlated with the preceding neuronal properties (9).…”

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confidence: 95%

Growth of a rat neuroblastoma cell line in serum-free supplemented medium.

Bottenstein¹,

Sato²

1979

Proc. Natl. Acad. Sci. U.S.A.

1,944

983

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The rat neuroblastoma B104 cell line, which originated in the central nervous system, was able to proliferate in the absence of serum in synthetic medium supplemented with insulin, transferrin, progesterone, selenium, and putrescine. When added individually, each supplement had little or no effect; however, in combination there was a marked synergistic effect on cell number. The cells attained the same saturation density in this medium as in medium with 10% fetal calf serum added. More extensive process formation was observed in the supplemented medium, and other differentiated properties were retained as well. Synthetic media generally require serum supplementation to support the proliferation or survival of cultured cells. The inclusion of serum, however, may significantly affect experimental reproducibility, because batch variations occur as a result of differences in donor age, sex, nutrition, and physiological state even in pooled serum samples. In addition, the complex undefined nature of serum is a complication when assessing the effect(s) of regulatory agents, such as hormones or neurotransmitters, on differentiated properties of nervous system cells in culture. This is particularly important for long-term studies, because if serum is deleted a substantial reduction of cell numbers, and in many cases complete cell death, may occur within hours or a few days.In order to circumvent these problems, several cell lines have been adapted to proliferate in serum-free media (1-4). How The B104 rat neuroblastoma, a cell line of central nervous system origin, exhibits many of the properties characteristic of differentiated neurons, such as generation of action potentials, synthesis of neurotransmitters, and presence of neurotransmitter receptors and the neuron-specific 14-3-2 protein (9, 10). Furthermore, B104 cells, like C1300 neuroblastoma cells (11, 12), respond to removal of serum by rapidly extending neurites, a phenomenon that has been correlated with the preceding neuronal properties (9).In this communication we report that B104 cells can proliferate in a serum-free synthetic medium supplemented with insulin, transferrin, progesterone, selenium, and putrescine (N2 medium). More extensive process formation is seen in N2 than in serum-supplemented medium, and other differentiated properties have been retained as well. METHODS AND MATERIALSCell Culture. The rat neuroblastoma B104 cell line was obtained from D. Schubert of the Salk Institute, La Jolla, CA. Stock cultures were maintained in a 1:1 mixture of Ham's F12 medium and the Dulbecco-Vogt modification of Eagle's medium (DME) supplemented with 5% (vol/vol) horse serum; 2.5% (vol/vol) fetal calf serum; 1.2 g of NaHCO3 per liter; 15 mM Hepes buffer; and 40 mg of penicillin, 8 mg of ampicillin, and 90 mg of streptomycin per liter. Triple-distilled water was used to prepare media. Stock cultures were grown in flasks (Falcon Plastics 3013, 25-cm2 surface area) in 4 ml of medium in a humidified atmosphere of 5% C02/95% air at 370C and subcultured every ...

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“…This suggests that approaches to cancer therapy should be sought that do not involve cell kill; one such approach envisions the conversion of malignant cells through induced differentiation to benign forms with no proliferative potential. This approach assumes that malignancy is not an irreversible state, as has been demonstrated with a variety of tumour types, including teratocarcinomas (Pierce & Wallace, 1971;Brinster, 1974), neuroblastomas (Schubert et al, 1971;Prasad, 1983) squamous cell carcinomas (Pierce & Wallace, 1971), leukaemias (Metcalf et al, 1969;Paran et al, 1970;Gootwine et al, 1982), and adenocarcinomas of the breast (Decosse et al, 1973;Pierce et al, 1977). The use of differentiation as a therapeutic approach also presumes that cancer is a disease of altered maturation.…”

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confidence: 99%

The 1985 Walter Hubert lecture. Malignant cell differentiation as a potential therapeutic approach

Sartorelli¹

1985

Br J Cancer

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it was possible to demonstrate that physiological levels of retinoic acid and epidermal growth factor were capable of preventing the differentiation of these malignant keratinocytes into a mature tissue-like structure. The terminal differentiation caused by certain antineoplastic agents was investigated in HL-60 promyelocytic leukaemia cells to provide information on the mechanism by which chemotherapeutic agents induce cells to by-pass a maturation block. The anthracyclines aclacinomycin A and marcellomycin were potent inhibitors of N-glycosidically linked glycoprotein biosynthesis and transferrin receptor activity, and active inducers of maturation; temporal studies suggested that the biochemical effects were associated with the differentiation process. 6-Thioguanine produced cytotoxicity in parental cells by forming analog nucleotide. In hypoxanthine-guanine phosphoribosyltransferase negative HL-60 cells the 6-thiopurine initiated maturation; this action was due to the free base (and possibly the deoxyribonucleoside), a finding which separated termination of proliferation due to cytotoxicity from that caused by maturation.

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Induced differentiation of a neuroblastoma

Cited by 226 publications

References 5 publications

Maturation of neuroblastoma cells in the presence of dimethylsulfoxide.

Maturation of neuroblastoma cells in the presence of dimethylsulfoxide.

Growth of a rat neuroblastoma cell line in serum-free supplemented medium.

The 1985 Walter Hubert lecture. Malignant cell differentiation as a potential therapeutic approach

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