Growth Activation of Cell Colonies in vitro by Extracts of Adult Tissues

Hoffman, R. S.; Tenenbaum, E.; Doljanski, L.

doi:10.1038/143764c0

Cited by 8 publications

(4 citation statements)

References 3 publications

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“…The in vitro counterpart of this experiment was conducted in 1939 (2)(3)(4), showing the mitogenic effect of brain homogenates on cultured fibroblasts. However, it was not until 1972t that an autoregulatory role of brain-derived growth factors on brain cells was first demonstrated in this laboratory (5,6).…”

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confidence: 99%

Purification and characterization of glia maturation factor beta: a growth regulator for neurons and glia.

Lim

Miller

Zaheer

1989

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

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A protein has been isolated from bovine brains by using a modification of the procedure used to purify glia maturation factor. The method consists of ammonium sulfate precipitation, chromatography with DEAE-Sephacel, Sephadex G-75, and hydroxylapatite columns, passage through a heparin-Sepharose column, and rinally fractionation by reverse-phase HPLC with a C4 column. The isolated protein reacts strongly with the mouse monoclonal antibody G2-09 and has a molecular weight of =17,000 and an isoelectric point of pH 4.9. The N terminus is blocked, but tryptic digestion releases 28 peptides, 8 of which have been sequenced. The total known residues add up to more than two-thirds of the entire 140-residue protein, estimated from amino acid composition, and show no sequence homology with any known protein.Reversible thermal renaturation greatly enhances its biological activity. The purified protein stimulates differentiation of normal neurons as well as glial cells. It inhibits the proliferation of the N-18 neuroblastoma line and the C6 glioma line while promoting their phenotypic expression. We designate this protein glia maturation factor f.The trophic function of the nervous system on peripheral organs was detected in an in vivo system as early as 1823 (1). The in vitro counterpart of this experiment was conducted in 1939 (2-4), showing the mitogenic effect of brain homogenates on cultured fibroblasts. However, it was not until 1972t that an autoregulatory role of brain-derived growth factors on brain cells was first demonstrated in this laboratory (5,6). This factor was named glia maturation factor (GMF), based on the bioassay system in which the activity was first observed. Since then this laboratory has engaged in an intensive search for the molecule responsible for this important function. The effort culminated in the isolation of a brain protein with a unique amino acid sequence documented below. MATERIALS AND METHODSPreliminary Purification of GMF-j3. The published procedure (7, 8) through the Sephadex G-75 step was followed with slight modifications. Briefly, four beef brains [1.0 kg (total wet weight)] were homogenized and centrifuged to obtain the crude extract. The ammonium sulfate precipitate between 45% and 70% saturation was dissolved in 100 ml of water and dialyzed for two 6-hr periods against 10 liters of water. The sample was adjusted to contain 0.02 M Tris HCl (pH 7.45) and applied to a DEAE-Sephacel column (2.5 x 37 cm). After eluting with 1.25 liters of a linear gradient of 0-0.3 M NaCl in the same buffer at 50 ml/hr, the fractions that showed mitogenic and morphologic activities on astrocytes were pooled (500 ml) and concentrated to 50 ml by Amicon PM10 filtration. The sample was applied to a Sephadex G-75 column (5 x 90 cm) and eluted with 0.15 M NaCl containing the above buffer at 40 ml/hr. The fractions active on astrocytes were pooled (500 ml) and used for the final steps of purification (see Results).Bioassay Production of Antibodies and Immunoassay. The mouse monoclonal antibody G2-0...

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confidence: 99%

Purification and characterization of glia maturation factor beta: a growth regulator for neurons and glia.

Lim

Miller

Zaheer

1989

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

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“…More recently, however, a group at the Hebrew University at Jerusalem has made studies leading them to believe that adult-tissue extracts are as good, if not better, than embryo extracts for the promotion of culture growth. Hoffman, Tenenbaum & Doljanski (1938) found that extracts from adult heart, smooth muscle, striated muscle, brain, lung, liver, kidney, testes, ovary, oviduct and pancreas were all more effective than extracts from comparable embryonic structures, with the extracts from such non-mitotic tissues as adult heart and brain giving culture areas some two to three times greater than the corresponding EE preparations. respectively for cultures in the three systems.…”

Section: Adult-tissue Extractsmentioning

confidence: 95%

“…respectively for cultures in the three systems. Hoffman, Tenenbaum & Doljanski (1938) found that extracts from adult heart, smooth muscle, striated muscle, brain, lung, liver, kidney, testes, ovary, oviduct and pancreas were all more effective than extracts from comparable embryonic structures, with the extracts from such non-mitotic tissues as adult heart and brain giving culture areas some two to three times greater than the corresponding EE preparations. Apparently a species effect exists, however, since Doljanski, Hoffman & Tenenbaum (1942) found that extracts from the hearts of adult sheep, cows, rabbits and dogs showed comparable effects when tested on chick heart fibroblasts, whereas Doljanski et al (1944) found that similar extracts from hamsters and rats were somewhat inhibitory under the same conditions.…”

Section: Adult-tissue Extractsmentioning

confidence: 95%

The Liquid Medium in Tissue Culture

Stewart

Kirk

1954

Biological Reviews

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Summary In the past, largely on an empirical basis, it was found that the most effective liquid media for culturing animal tissue cells consisted of mixtures of physiological saline solutions, tissue extracts (preferably from embryonic tissue) and serum or plasma. The marked influx of investigators with primarily biochemical interests into the tissue culture field in the last few decades has led to determined efforts to learn more of the precise relationships between the composition of these media and the metabolism of the tissues. Since both of these are so complex, and the equilibrium between them is such a delicate one, it is perhaps not surprising that many apparently contradictory results are seen in the current literature. The place of physiological salt solutions in tissue cultures appears to be a relatively simple matter, and their functions are reasonably well understood. The present confusion relates to the role of the tissue extract and serum components of the medium. For certain types of cells there is evidence that a tissue‐extract nucleo‐protein fraction, supplemented by a surprisingly small group of smaller molecule nutrients, is adequate to support growth, at least for short periods. Various possible mixtures of these smaller components may be effective as a supplement in the same system, although it is probable that optimum combinations exist, again depending on the cell type and the culturing conditions. The tissue nucleoprotein fraction cannot be considered as a ‘growth hormone’, in the conventional sense of that term, since other cell types, or even superficially similar cell types taken from other animal donors, are able to maintain themselves satisfactorily when this fraction is completely absent, at least if serum proteins are present in the medium. It is thus evident that quantitative comparisons of data found by various researchers are of very dubious validity unless it is certain that identical culturing techniques were applied, using identical cell systems. On a more positive note, however, it seems probable that the analytical approach to the liquid medium problem is bringing the long‐sought goal of a controlled composition ‘synthetic’ medium for cultured animal cells much nearer to attainment for certain systems sharply defined as to cell type and culturing procedures. There are also indications that ‘growth’, biochemically speaking, as defined by the cell division in a culture, may be a separate process from ‘growth’ as defined by increase in culture mass, protein content or similar criteria. In other words, the chemical and physical conditions required to aid in inducing or in completing the mitotic cycle are not necessarily directly related to the nutritional requirements of growing and maturing individual cells in a healthy culture. There is some evidence that extracts made from tissues of older animals are, in many culture systems, as effective as extracts of embryonic tissue. Since, on the whole, such tissues are more accessible, their wider use by tissue culturists should be in...

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“…From these experiments little could be concluded as to a possible correlation of growth promoting activity and the state of proliferation and differentiation of the donor tissue since whole embryo extracts include components of tissues at widely differing stapes of development. Examination of extracts from adult tissues with different rates of cell proliferation did not indicate that proliferative activity in a tissue and growth promoting activity of its extracts are related ( 2 ) . -4s a result of a more systematic study of the cytological and chemical changes in developing leg muscle of the chick ( 3 , 4 ) , data have become available which invite a reconsideration of the relationships of the developmental stage in a single tissue and the growth promoting activity of its extracts.…”

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confidence: 98%