scholarly journals STUDIES UPON THE CHARACTERISTICS OF DIFFERENT CULTURE MEDIA AND THEIR INFLUENCE UPON THE GROWTH OF TISSUE OUTSIDE OF THE ORGANISM

1912 ◽  
Vol 16 (4) ◽  
pp. 421-431 ◽  
Author(s):  
Ragnvald Ingebrigtsen
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Connective Tissue ◽  
Culture Medium ◽  
Culture Media ◽  
Embryonic Tissue ◽  
Adult Tissue ◽  
Ringer’S Solution ◽  
Tissue Cells

1. There is a great difference between embryonic and adult tissue as far as their growth outside of the organism is concerned. Adult tissue grows only in plasma. Embryonic tissue grows also very well in serum and serum plus agar. In Ringer's solution and in Ringer's solution plus agar no growth occurs, whether embryonic or adult tissue is employed; survival and emigration of cells are seen to some extent. 2. For the growth of connective tissue cells of chick embryo, unheated homogenic serum is a better culture medium than heated serum. The growth of epithelial cells is not thus influenced. 3. Heated heterogenic serum is a better culture medium for growth of embryonic connective tissue cells than unheated. 4. There is an inverse ratio between the hemolytic power of heterogenic sera and the extent of growth of tissue in them. This inverse ratio is not found in heterogenic plasmas.

scholarly journals CONTRIBUTIONS TO THE STUDY OF THE MECHANISM OF THE GROWTH OF CONNECTIVE TISSUE

1913 ◽  
Vol 18 (3) ◽  
pp. 287-298 ◽  
Author(s):  
Alexis Carrel
Keyword(s):  
Connective Tissue ◽  
Culture Medium ◽  
Dynamic Condition ◽  
Real Life ◽  
Rate Of Growth ◽  
Independent Life ◽  
Tissue Cells ◽  
Chick Embryo Heart ◽  
Embryo Heart

When connective tissue cells have been cultivated for a certain length of time in a medium which has been repeatedly changed, a definite relation arises between the rate of growth of the cells and the composition of the medium. It is possible, by adding to the culture medium a given quantity of certain substances, such as embryonic juices, to foresee the extent to which a fragment of tissue composed of a given strain of cells will increase in a given time. The rate of growth of a strain of cells can be accelerated or retarded by the addition to the medium of activating or retarding substances. The dynamic condition of a strain of connective tissue cells, which have been living in a given medium for some time, is not a definitely acquired characteristic, but a temporary state, and is the product or function of the medium in which the cells are living, and is readily modified merely by altering the composition of the medium. A knowledge of the characteristics of the growth of connective tissue described has led to a new result,—the indefinite proliferation of a strain of connective tissue cells outside of the organism. The strain of connective tissue originally obtained from a fragment of chick embryo heart, which had been pulsating in vitro for 104 days, was still actively alive after sixteen months of independent life and more than 190 passages. The rate of proliferation of the connective tissue sixteen months old equalled and even exceeded that of fresh connective tissue taken from an eight day old embryo. It appears, therefore, that time has no effect on the tissues isolated from the organism and preserved by means of the technique described above. During the sixteenth month of life in vitro the cells increased rapidly in number and were able in a short time to produce a large quantity of new tissue. This fact, therefore, definitely demonstrates that the tissues were not in a state of survival, as was the case in certain earlier experiments, but in a condition of real life, since the cells of which they were composed, like microorganisms, multiplied indefinitely in the culture medium.

scholarly journals ACTION ON FIBROBLASTS OF EXTRACTS OF HOMOLOGOUS AND HETEROLOGOUS TISSUES

1923 ◽  
Vol 38 (5) ◽  
pp. 499-511 ◽  
Author(s):  
Alexis Carrel ◽  
Albert H. Ebeling
Keyword(s):  
Chick Embryo ◽  
Guinea Pigs ◽  
Embryonic Tissue ◽  
Rabbit Serum ◽  
Cell Multiplication ◽  
Tyrode Solution ◽  
Adult Tissue ◽  
Tissue Extracts ◽  
Adult Tissues ◽  
Pure Cultures

Extracts of homologous adult tissues detemine an increase in the mass of pure cultures of chicken fibroblasts nourished thereon comparable to that resulting from embryonic tissue juice. But the effect of these extracts differs markedly from that of the latter, since cell multiplication does not continue indefinitely. After a few passages, the fibroblasts cultivated in adult tissue extracts grew more slowly than in Tyrode solution. The cytoplasm became dark and filled with fat granules, and death followed. It is possible that the tissues of adult animals contain, as does the serum, substances which are toxic for the homologous cells, and which progressively overcome the effect of the growth-activating substances. The effect of heterologous adult tissue extracts did not differ markedly from that of homologous tissues. The chicken connective tissue increased slightly in mass, but died sooner than the controls in Tyrode solution. By contrast, tissue juices derived from the embryos of mice, guinea pigs, and rabbits acted on chicken fibroblasts in the same manner as chick embryo juices. The increase in mass of the cultures was regular and rapid. They doubled in size every 48 or 72 hours, and the rate of growth did not decrease after 30 days. It appears that embryonic tissue juices are not necessarily toxic for heterologous fibroblasts, and that they can be used in the building up of protoplasm in the tissues of a different species. In experiments made long ago, the action of tissue juice was described as being specific. The premature death of the fibroblasts cultivated in heterologous juices at that time would now appear to have been due to spontaneous changes in the pH and the deterioration that even normal chick embryo juice at a pH of 7.8 undergoes spontaneously. In the recent experiments, when freshly prepared homologous and heterologous juices were used, their action on chicken fibroblasts in pure culture was identical. However, the fibroblasts produced in cultures nourished by rabbit juice grew better when transferred to rabbit serum than did ordinary chicken fibroblasts. It has not been determined as yet whether this effect is due to an immunization of the fibroblasts against rabbit humors, or to some decrease in the specificity eventuating in cells intermediate between rabbit and chicken fibroblasts. It may be concluded that, under the conditions of the experiments : 1. Pure cultures of chicken fibroblasts increase in mass under the influence of extracts of adult homologous tissues. But they ultimately die while the fibroblasts cultivated in embryonic tissue juices live indefinitely. 2. The increase in mass of chicken fibroblasts cultivated in the juices of mouse, guinea pig, rabbit, and chick embryos is about identical. 3. Chicken fibroblasts produced in cultures nourished by rabbit embryonic tissue juice are less sensitive to the inhibiting action of rabbit serum than ordinary chicken fibroblasts. 4. Cultures of chicken fibroblasts in extracts of adult tissues of mice, guinea pigs, and rabbits increase slightly in mass, but the increase is temporary and death occurs after a few passages.

scholarly journals Differences in the growth of transplantable tumours in plasma and serum culture media

1933 ◽  
Vol 112 (776) ◽  
pp. 250-263 ◽  
Keyword(s):  
Connective Tissue ◽  
Culture Medium ◽  
Culture Media ◽  
Vital Staining ◽  
Fundamental Difference ◽  
Tissue Cultures ◽  
Malignant Cells ◽  
Cover Glass ◽  
Controlled Conditions ◽  
Fibrin Network

A recent study of the behaviour of normal and malignant cells in tissue cultures of transplantable tumours (Ludford, 1932, c ), carried out by means of vital staining has revealed a fundamental difference between them. When a plasma culture undergoes liquefaction the outgrowth of malignant cells collapses as the result of the breaking down of the fibrin network by which the cells are supported. Malignant cells round off, usually become detached, degenerate and die. Non-malignant connective-tissue cells remain adhere to the cover-glass, and without renewal of the culture medium, often survive after all the malignant cells have diet. Such behaviore suggest a difference between the plasma membrance of mormal and malignas cells would appear to be lees adihesive. On the basis of these observation it appeared improbable that any outgrowth of maligbant cells could occur in a serum medium. Since conclusion of far-reaching Important have been drawn from experiments with culture of tumours, it seemed desirable to investigate matter under controlled conditions.

scholarly journals On the variation in the growth of mammalian tissue in vitro according to the age of the animal

1915 ◽  
Vol 88 (606) ◽  
pp. 476-482
Keyword(s):  
Culture Medium ◽  
Embryonic Tissue ◽  
Mammalian Tissue ◽  
Adult Tissue ◽  
Suitable Medium ◽  
Previous Communication ◽  
Series Of Experiments

In a previous communication it was shown that there was considerable variation in the value, as a culture medium, of plasmata taken from different animals of the same species; that these plasmata did not vary as to whether they were homogenous or autogenous, but that some plasmata were good media and some were bad. During this investigation certain evidence arose that this difference might in part be due to the age of the animal. In the present investigation a series of experiments was carried out to show what was the effect, if any, of the age of the animal upon the plasma as culture medium, and upon the tissues as regards the power of growth. Carrel, Burrows, Harrison, and Ingebrigtsen have shown in several papers that embryonic tissue tends to grow more rapidly and more vigorously than adult tissue. There appears to have been, however, no work conducted on the characters of the plasma, although it has been frequently assumed that the plasma of the young or embryonic animals makes a more suitable medium than that of adults; nevertheless it was permissible to believe that the reverse might in fact be true, and that the plasma of young animals is a less suitable medium. It would appear important that this point should be settled, that thereby evidence might be gained as to the controlling influences on the growth of young tissue in vivo .

scholarly journals A STRAIN OF CONNECTIVE TISSUE SEVEN YEARS OLD

1919 ◽  
Vol 30 (6) ◽  
pp. 531-537 ◽  
Author(s):  
Albert H. Ebeling
Keyword(s):  
Connective Tissue ◽  
Culture Medium ◽  
Rate Of Growth ◽  
Tissue Cells

1. A strain of connective tissue is still very active after more than 7 years of life in vitro. 2. The rate of growth of the fragments of tissue can be measured accurately and used for testing the action of many different factors on the growth of connective tissue cells. 3. The rate of growth of the strain is at least as rapid as it was 5 years ago, and may be more active. 4. The connective tissue cells appear to have the power of multiplying indefinitely in a culture medium, as do microorganisms.

scholarly journals Heavy meromyosin labeling of intermediate filaments in cultured connective tissue cells.

1978 ◽  
Vol 78 (3) ◽  
pp. 644-652 ◽  
Author(s):  
I K Buckley ◽  
T R Raju ◽  
M Stewart
Keyword(s):  
Chick Embryo ◽  
Connective Tissue ◽  
Intermediate Filaments ◽  
Radical Change ◽  
Accessory Proteins ◽  
Heavy Meromyosin ◽  
Binding Properties ◽  
Intermediate Size ◽  
Tissue Cells ◽  
In Cells

Mild treatment with trypsin causes a radical change in the heavy meromyosin (HMM) binding properties of intermediate filaments in glycerinated, myosin-extracted cultured chick embryo connective tissue cells. In non-trypsin-treated cells, HMM labeling of filaments was often indistinct and variable in its distribution. By contrast, in cells treated with trypsin (under conditions which allowed most intermediate filaments to survive), virtually all filaments, including those of intermediate size, decorated with HMM to give distinct arrowhead patterns. We suggest that most intermediate filaments in such cells contain a core of F-actin masked by trypsin-labile accessory proteins.

Human biliary mucin binds to E-selectin: a possible role in modulation of inflammation

2001 ◽  
Vol 280 (5) ◽  
pp. G1043-G1048 ◽  
Author(s):  
Nair Sreejayan ◽  
Bianca M. Wittig ◽  
Niko von Stillfried ◽  
Matthias S. Hennicke ◽  
Gunther Meyer ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Culture Medium ◽  
Culture Media ◽  
Binding Activity ◽  
Side Chain ◽  
Human Gallbladder ◽  
Human Bile ◽  
Sialyl Lewis ◽  
Selectin Binding ◽  
Bile Samples

E-selectin, expressed on endothelial cells, mediates adhesion of leukocytes and tumor cells to endothelium. CA19-9 (sialyl-Lewisa) and sialyl-Lewisx are specific ligands for E-selectin. We have recently shown that mucin-rich culture media from human gallbladder epithelial cells contains CA19-9. In this study, we have tested whether human biliary mucin binds to E-selectin. The ability of mucins to inhibit the adhesion of HL-60 cells to immobilized E-selectin was taken as an index for E-selectin binding. Gallbladder bile, hepatic bile, and culture medium from human gallbladder epithelial cells completely inhibited the adhesion of HL-60 cells to E-selectin. The mucin-rich fractions of human bile exhibited strong inhibition, whereas mucin-free fractions had little effect. In contrast to human bile samples, CA19-9-free medium from cultured dog gallbladder epithelial cells failed to inhibit HL-60 binding. Furthermore, after CA19-9 immunoaffinity chromatography, which selectively extracted CA19-9 from bile, bile samples showed poor inhibition of HL-60 adhesion to immobilized E-selectin. A good correlation was observed between E-selectin binding and CA 19-9 concentrations in bile. Our results show that human bile has E-selectin binding activity that is mediated by the CA19-9 side chain of biliary mucin.

scholarly journals CULTIVATION OF THE SKIN EPITHELIUM OF THE ADULT FROG, RANA PIPIENS

1914 ◽  
Vol 20 (6) ◽  
pp. 614-634 ◽  
Author(s):  
Eduard Uhlenhuth
Keyword(s):  
Epithelial Cells ◽  
Connective Tissue ◽  
Basal Layer ◽  
Vertical Direction ◽  
Middle Layer ◽  
Leopard Frog ◽  
Nutritive Medium ◽  
Physicochemical Conditions ◽  
Tissue Cells

1. Fragments of skin taken from the back of the leopard frog were cultivated in a mixture of plasma and muscle extract of the same species. 2. A few hours after explanation, processes of activity are seen to arise, which finally lead to the formation of a compact epithelial rim around the fragment of skin. 3. These epithelial cells undergo gradual transformation into a spindle-shaped type of cell; in this form they resemble the spindle cells which have been described as connective tissue cells. 4. The growth of a rim of tissue around the explanted fragments of skin may be ascribed principally to the activity of a basal layer of epithelial cells, the units of which first advance into the medium as compact membranes by means of the so called epithelial movement, but which later become separated from the compact membrane, and having assume a spindle form, spread according to the manner of connective tissue cells. 5. In contradistinction to this basal layer, the cells of an upper and middle layer of epithelium upon separation from the fragment of skin and isolation in the nutritive medium, remain completely inactive. 6. The connective tissue of frog skin, for reasons not yet completely established, does not participate in the production of the rim of cells, possibly for the reason that it was early surrounded by the epithelium and was thus prevented from sending out cells into the medium. 7. The transformation of the epithelial cells into a type of cells of the spindle form is a result of the changes in the physicochemical conditions brought about by the life in vitro, which become similar to the physicochemical conditions normally characteristic of connective tissue. 8. The final stage of the explanted fragments of skin is a hollow epithelial sphere (cystic stage) filled with connective tissue. 9. During the cystic stage the epithelium again shows its normal polar differentiation, as a consequence of the physicochemical conditions which now approach normal. 10. These conditions also permit of the shedding of cells in a vertical direction, although previously the production of cells in a horizontal direction had already become impossible.

scholarly journals Dispersed mammary gland epithelial cells. I. Isolation and separation procedures.

1977 ◽  
Vol 72 (2) ◽  
pp. 390-405 ◽  
Author(s):  
J P Kraehenbuhl
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Connective Tissue ◽  
Structural Integrity ◽  
Plasma Cells ◽  
Buoyant Density ◽  
Light And Electron Microscopy ◽  
Enzymatic Digestion ◽  
Myoepithelial Cells ◽  
Tissue Cells

The mammary gland from midpregnant rabbits has been dissociated into individual cells by enzymatic digestion, divalent cation chelation, and gentle shearing. A heterogeneous cell population is obtained, comprising approximately 60% parenchymal cells, approximately 10% myoepithelial cells, and approximately 30% connective tissue cells, including fibroblasts, plasma cells, and microphages. The epithelial cells are characterized by the presence of fat droplets, which in 65% of the cells form large supranuclear vacuoles. Their buoyant density is less than 1.045, allowing their separation from myoepithelial cells and connective tissue cells by isopycnic centrifugation in a density gradient. The homogeneity of the epithelial cell fraction has been assessed by light and electron microscopy. The cells are viable and functionally active as indicated by their ability to exclude vital dyes, incorporate labeled precursors, consume oxygen, maintain intracellular Na+ and K+ concentrations, and retain their structural integrity. In addition, when cultured in Petri dishes, the cells grow as a monolayer, reestablish junctional complexes and retain cell polarity.

Factors Involved In the Plasminogen Activation System in Human Breast Tumours

1994 ◽  
Vol 71 (05) ◽  
pp. 684-691 ◽  
Author(s):  
László Damjanovich ◽  
Csaba Turzó ◽  
Róza Ádány
Keyword(s):  
Epithelial Cells ◽  
Connective Tissue ◽  
Plasminogen Activators ◽  
Breast Tumour ◽  
Plasminogen Activation ◽  
Malignant Tumours ◽  
Plasminogen Activation System ◽  
Activation System ◽  
Malignant Breast ◽  
Pai 1

SummaryThe plasminogen activation system is a delicately balanced assembly of enzymes which seems to have primary influence on tumour progression. The conversion of plasminogen into serine protease plasmin with fibrinolytic activity depends on the actual balance between plasminogen activators (urokinase type; u-PA and tissue type; t-PA) and their inhibitors (type 1 and 2 plasminogen activator inhibitors; PAI-1 and PAI-2). The purpose of this study was to determine the exact histological localization of all the major factors involved in plasminogen activation, and activation inhibition (plasmin system) in benign and malignant breast tumour samples. Our results show that factors of the plasmin system are present both in benign and malignant tumours. Cancer cells strongly labelled for both u-PA and t-PA, but epithelial cells of fibroadenoma samples were also stained for plasminogen activators at least as intensively as tumour cells in cancerous tissues. In fibroadenomas, all the epithelial cells were labelled for PAM. Staining became sporadic in malignant tumours, cells located at the periphery of tumour cell clusters regularly did not show reaction for PAI-1. In the benign tumour samples the perialveolar connective tissue stroma contained a lot of PAI-1 positive cells, showing characteristics of fibroblasts; but their number was strongly decreased in the stroma of malignant tumours. These findings indicate that the higher level of u-PA antigen, detected in malignant breast tumour samples by biochemical techniques, does not necessarily indicate increased u-PA production by tumour cells but it might be owing to the increased number of cells producing u-PA as well. In malignant tumours PAI-1 seems to be decreased in the frontage of malignant cell invasion; i.e. malignant cells at the host/tumour interface do not express PAI-1 in morphologically detectable quantity and in the peritumoural connective tissue the number of fibroblasts containing PAI-1 is also decreased.

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