Selective cell culture of brain cells by serum-free, hormone-supplemented media: A comparative morphological study

Brunner, Gerd; Lang, K.; Wolfe, Richard A.; McClure, Don B.; Sato, Gordon

doi:10.1016/0165-3806(81)90024-9

Cited by 62 publications

(24 citation statements)

References 20 publications

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“…The replacement of serum as a supplement to culture medium provides a number of advantages (1,(23)(24)(25)(26), including elimination from the in vitro environment of growth inhibitors in the serum, allowing culture of cell types that otherwise could not be cultured on a long-term basis. Another approach that has been successful for extending cell growth in vitro is the supplementation of serum-containing culture medium with a high concentration of albumin, approximating the protein concentration of plasma and interstitial fluid (34).…”

Section: Discussionmentioning

confidence: 99%

“…3T3 cells (7) were cultured in medium supplemented with 10% calf serum. For the culture of adult mouse cells, brains of 6-week-old BALB/c mice were dissociated as described (26), washed with medium, and cultured as described for SFME cells, with additional supplementation of fibroblast growth factor (FGF) at 10 ng/ml and heparin at 1 lug/ml (3 8379 from mouse brain, minced tissue fragments were treated as described above for cell suspensions.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Serum and transforming growth factor beta regulate glial fibrillary acidic protein in serum-free-derived mouse embryo cells.

Sakai

Rawson

Lindburg

et al. 1990

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

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Serum-free mouse embryo (SFME) cells, derived in medium in which serum is replaced with growth factors and other supplements, display distinctive properties: (i) SFME cells do not lose proliferative potential or show gross chromosomal aberration upon extended culture, (ii) these cells depend on epidermal growth factor for survival; and (ii) SFME cell proliferation is reversibly inhibited by serum. Treatment of SFME cells with serum or transforming growth factor j3 led to the appearance of glial fibrillary acidic protein, a specific marker for astrocytes. The appearance of glial fibrillary acidic protein in cultures was reversed upon removal of transforming growth factor (3 or serum. Cells with properties similar to SFME cells were also isolated from adult mouse brain. These results suggest a role for transforming growth factor (3 in astrocyte differentiation in developing organisms and in response to injury and identify the cell type that has the unusual properties of SFME cells.Serum-free mouse embryo (SFME) cells initiated and maintained multipassage in a rich basal nutrient medium supplemented with insulin, transferrin, epidermal growth factor (EGF), high-density lipoprotein, and fibronectin display several distinctive properties (1)(2)(3)(4)(5)(6). Mouse embryo cells derived in conventional serum-supplemented medium undergo a period in which proliferative potential is lost or reduced, and then genetically altered cells with indefinite proliferative potential emerge (2,7,8). SFME cells do not exhibit any loss of proliferative potential or gross chromosomal aberration and maintain a relatively stable karyotype when cultured for >10 times the population doublings achieved with karyotypically normal mouse embryo cells in serum-containing medium. Although the period in which reduced growth rate can be detected may be minimal or absent in cultures in serumcontaining medium maintained in a passaging protocol involving high plating densities, the protocols used to derive SFME cells lead to a well-defined period of limited proliferative potential in cultures maintained in serum-containing medium (2, 7) that is not seen when the cells are cultured in serum-free medium. SFME cells also depend on EGF for survival, and cycloheximide or actinomycin D prevents death from EGF deprivation (5), suggesting that cell death requires the synthesis of RNA and protein, a phenomenon of programmed cell death similar to that reported for neuronal cell death in the absence of nerve growth factor (NGF) (9). In addition, proliferation of SFME cells is reversibly inhibited by serum or platelet-free plasma (1, 4, 6), and thus these cells would not be propagated under conventional culture conditions using serum-supplemented medium. As for most mouse embryo cell lines, the precise nature of the cells giving rise to the continuously growing SFME cultures was unknown. The unusual characteristics of SFME cells led us to attempt to determine the cell type represented by these cells.We found that treatment of SFME cells with serum or We a...

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Serum and transforming growth factor beta regulate glial fibrillary acidic protein in serum-free-derived mouse embryo cells.

Sakai

Rawson

Lindburg

et al. 1990

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

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“…Since the pioneering work by the group of Sato (see Barnes and Sato, 1980), serum-free, hormone-supplemented culture media have been used also for the culture of neural cells (Honneger et al, 1979;Snyder and Kim, 1979;Morrison and de Vellis, 1981;Romijn et al, 1982;Brunner et al, 1982). One of the advantages of a chemically defined medium over a serum-containing medium is the possibility to study quantitatively and reproducibly the influence of hormones upon myelination.…”

Section: Discussionmentioning

confidence: 99%

“…Serum-free media for primary cultures of neural cells have been used previously (e.g. Honneger et al, 1979;Kim, 1979: Morrison andde Vellis, 1981;Romijn et al, 1982;Brunner et al, 1982). This paper describes a method for the isolation of an oligodendrocyte-enriched cell population from the brains of rat pups.…”

Section: Introductionmentioning

confidence: 99%

Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

Koper

Lopes‐Cardozo

Romijn

et al. 1984

Journal of Neuroscience Methods

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Kew'ords: oligodendrocyte--primary culture--serum-free medium--chemically defined medium--brain cells--rat brain Oligodendrocytes were isolated from the cerebra of young rats (5-10 days old) by trypsinization of the tissue followed by cell separation on Percoll gradients. The isolation was carried out in physiological, isotonic media. The cell yield was 2-4 × 106 cells per brain; the plating efficiency was >/70%. Isolated cells were seeded on poly-L-lysine-coated culture dishes and maintained in a serum-free, chemically defined medium for at least 30 days. After 10 days in culture 67+10% of the surviving cells were oligodendrocytes, as judged by immunocytochemical and morphological criteria, whereas most of the other cells reacted positively with antiserum against glial fibrillary acidic protein. The expression of typical oligodendrocyte markers (2':3'-cyclic-nucleotide 3'-phosphodiesterase, galactocerebrosides and myelin basic protein) was greatly enhanced under these serum-free conditions as compared with cultures in serum-containing medium. The antigenic markers (galactocerebrosides, myelin basic protein) were absent in the freshly isolated cells but could be detected after 3 days in culture by immunocytochemistry. The activity of 2':3'-cyclic-nucleotide 3'-phosphodiesterase increased from 75 nmol min 1 mg 1 protein on day 4 to 400 nmol min-1 mg 1 protein on day 14 in culture.

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“…IA). Serum-free, supplemented medium was used to suppress glia and fibroblast growth (Brunner et al, 1982). The medium consisted of D-MEMIF12 (Life Technology) supplemented with 5 mg/ml insulin (Sigma, St. Louis, MO), 100 mg/ml transferrin (Sigma), 20 nM progesterone (Sigma), 100 mM putrescene (Sigma), 20 nM hydrocortisone (Sigma), 30 nM selenium (Sigma), and 0.035 mg/ml albumin (Sigma).…”

Section: Fluorescent Labeling Of Thalamocortical Axonsmentioning

confidence: 99%

Addition of tetrodotoxin alters the morphology of thalamocortical axons in organotypic cocultures

Wilkemeyer

Angelides

1996

J. Neurosci. Res.

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Selective cell culture of brain cells by serum-free, hormone-supplemented media: A comparative morphological study

Cited by 62 publications

References 20 publications

Serum and transforming growth factor beta regulate glial fibrillary acidic protein in serum-free-derived mouse embryo cells.

Serum and transforming growth factor beta regulate glial fibrillary acidic protein in serum-free-derived mouse embryo cells.

Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

Addition of tetrodotoxin alters the morphology of thalamocortical axons in organotypic cocultures

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