[6] The growth of cells in serum-free hormone-supplemented media

Bottenstein, Jane E.; Hayashi, Izumi; Hutchings, Sharon E.; Masui, Hideo; Mather, Jennie P.; McClure, Don B.; Ohasa, Sugayuki; Rizzino, Angie; Sato, Gordon; Serrero, Ginette; Wolfe, Richard A.; Wu, Reen

doi:10.1016/s0076-6879(79)58127-0

Cited by 290 publications

(140 citation statements)

References 19 publications

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“…The growth requirements have been determined in a similar manner for other cell lines, and apparently there are unique sets of requirements for maximal growth of specific cell types (5)(6)(7)(8). Common constituents for these cell lines are insulin and transferrin, although the optimal concentrations may differ.…”

Section: Discussionmentioning

confidence: 99%

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

Bottenstein¹,

Sato²

1979

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

1,948

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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Section: Discussionmentioning

confidence: 99%

“…In order to circumvent these problems, several cell lines have been adapted to proliferate in serum-free media (1)(2)(3)(4) (5)(6)(7)(8).…”

mentioning

confidence: 99%

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

Bottenstein¹,

Sato²

1979

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

1,948

983

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“…We further demonstrated that soaking the pSi particles in chloroform prior to pressing caused localised etching of the PCL fibres, improving attachment of pSi microparticles, and that particle retention could further be improved by the deposition of a thin layer of PCL strands on top of the particles. Together, these improvements led to increased pSi particle loading compared with previous methods (Kashanian et al, 2010) h. We also demonstrated loading and release of a mixture of biologic agents containing epidermal growth factor, insulin and transferrin into the composite materials, using the proliferation of BALB/c 3T3 cells grown in serum-deprived medium (Bottenstein et al, 1979) as a readout. This functional assay confirmed that the biologics were released from the composites in an active form.…”

Section: Discussionmentioning

confidence: 87%

“…Test composites were incubated in 500 µl of 1 mg/ml insulin, 0.55 mg/ml transferrin, 0.67 µg/ml sodium selenite and 10 µg/ml EGF for 16 h at 4°C in a humidified box, to maintain activity of the biologics. Insulin, transferrin, and sodium selenite improve cell growth in serum-free or serum-low medium (Bottenstein et al, 1979). Composites incubated in PBS were used as controls.…”

Section: Loading and Release Of Insulin Transferrin Selenite And Egmentioning

confidence: 99%

A novel pressed porous silicon-polycaprolactone composite as a dual-purpose implant for the delivery of cells and drugs to the eye

Irani

Tian

Wang

et al. 2015

Experimental Eye Research

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Abbreviations: BSS ophthalmic balanced salt solution; DMEM Dulbecco's Modified Eagle's Medium; EGF epidermal growth factor; FBS fetal bovine serum; FDA fluorescein diacetate; PBS phosphate buffered saline; PCL polycaprolactone; pSi nanostructured porous silicon; SEM scanning electron microscopy; TEM transmission electron microscopy. To investigate loading of large-molecule biologics, murine BALB/c 3T3 cells, responsive to epidermal growth factor, insulin and transferrin, were seeded on composite materials. The cells showed significantly more proliferation at 48 h when seeded on composites loaded with these biologics, than on unloaded composites. No cell proliferation was observed on PCL alone, indicating the biologics had loaded into the pSi microparticles. Drug release, measured by ELISA for insulin, indicated a burst followed by a slower, continuous release over six days. When implanted under the rat conjunctiva, the most promising composite material did not cause significant neovascularization but did elicit a macrophage and mild foreign body

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“…These sera were chosen for their low content of corticosteroid: less than 5 ng/ml expressed in cortisol and monitored by radiocompetitive assays using human transcortine and they did not need further desteroidation before use. In some experiments, cells were grown in DH/4F medium [8] consisting of a mixture of DMEM and Ham-F12 (2: 1, v/v) supplemented with 5/~g/ml insulin, 5/~g/ml transferrin, 2/~g/ml luteinizing hormone and 10/~g/ml submaxillar gland extract. At the indicated period, cell cultures were washed twice with cold phosphate buffered saline, homogenized in 1 mM dithiothreitol, 25 mM Tris-HCI buffer (pH 7.5) in a Dounce homogenizer.…”

Section: Cell Culturementioning

confidence: 99%

Dexamethasone‐dependent expression of β^1–24 corticotropin stimulated adenylate cyclase during adipose conversion of 3T3‐F442A cells

Fève

Pairault

1987

FEBS Letters

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When 3T3-F442A preadipocytes were grown in culture media supplemented with corticosteroid poor fetal calf serum and insulin they differentiated into adipocytes. Glycerophosphate dehydrogenase, a marker of terminal differentiation, developed a 600-fold increase of activity whereas the adenylate cyclase system remained unresponsive to the synthetic ACTH(1-24) analog. In contrast, 3T3-F442A adipocytes, differentiated in the presence of dexamethasone, exhibited an adenylate cyclase activity which was stimulated 4-fold by ACTH(1-24). The stimulation of the adenylate cyclase activity by GTPyS remained unchanged (about 20-25-fold) suggesting that the G regulatory coupling protein was not functionally modified by dexamethasone. Binding studies with t25I-ACTH revealed that specific cellular binding could be evidenced in dexamethasone-treated cells while control adipocytes did not exhibit any specific binding of 125I-ACTH. These findings lend support to the hypothesis that the setting off of this ACTH responsiveness in 3T3-F442A cells is regulated by dexamethasone after cells are committed to adipose differentiation.

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[6] The growth of cells in serum-free hormone-supplemented media

Cited by 290 publications

References 19 publications

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

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

A novel pressed porous silicon-polycaprolactone composite as a dual-purpose implant for the delivery of cells and drugs to the eye

Dexamethasone‐dependent expression of β^1–24 corticotropin stimulated adenylate cyclase during adipose conversion of 3T3‐F442A cells

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[6] The growth of cells in serum-free hormone-supplemented media

Cited by 290 publications

References 19 publications

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

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

A novel pressed porous silicon-polycaprolactone composite as a dual-purpose implant for the delivery of cells and drugs to the eye

Dexamethasone‐dependent expression of β1–24 corticotropin stimulated adenylate cyclase during adipose conversion of 3T3‐F442A cells

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Dexamethasone‐dependent expression of β^1–24 corticotropin stimulated adenylate cyclase during adipose conversion of 3T3‐F442A cells