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Conditioned media

Fig. 3. Overview of puriftcation sequence for the nonrecombinant tissue plasminogen activator (t-PA) which also contains urokinase plasminogen activator (u-PA). Serum-free culture conditional media is from normal human ceU line. The temperature for aU. steps, except for size-exclusion chromatography... Fig. 3. Overview of puriftcation sequence for the nonrecombinant tissue plasminogen activator (t-PA) which also contains urokinase plasminogen activator (u-PA). Serum-free culture conditional media is from normal human ceU line. The temperature for aU. steps, except for size-exclusion chromatography...
Adsorption of t-PA to process equipment surfaces consisting of either stainless steel or glass was minimized by adding the detergent polyoxyethylene sorbitan monooleate (Tween 80) to the semm-free culture conditioned media at 0.01% (vol/vol). The equipment was also rinsed, before use, with phosphate buffered saline (PBS) containing 0.01% Tween 80. Hydrophilic, plastic equipment was used whenever possible. AH buffers were sterile filtered. Sterile filtration of Hquids and gases is usually carried out using 0.2 or 0.45 p.m filters. [Pg.46]

Treatment is carried out in 30% conditioned media. The serum concentration is 3% (3-h treatment) or 10% (treated more than 3 h). [Pg.213]

Tseng SC, Kruse FE, Merritt J, Li DQ. Comparison between serum-free and fibroblast-cocultured single-cell clonal culture systems Evidence showing that epithelial anti-apoptotic activity is present in 3T3 fibroblast-conditioned media. Curr Eye Res 15 973-984 (1996). [Pg.304]

Monocultures form the base of most cerebral endothelial cell culture models [105-107], In the case of filter-grown cells, some authors recommend coculture with astrocytes or the use of astrocyte-conditioned media [108-110], The necessity of this approach is presently under discussion, as cells which had been cultured without supplementation with conditioned media show similar features [94, 111],... [Pg.409]

The first experiments describing the clonal growth of hemopoietic progenitor cells immobilized in a soft gel matrix in vitro were reported by Bradley and Metcalf (1966) and Pluznick and Sachs (1965). Clonogenic cells are plated in the presence of various of feeder cells, medium conditioned by the growth of different tissues or cell lines in cultures (for example, 5637 bladder carcinoma conditioned media) and colony stimulating factors such as GM-CSF, G-CSF, M-CSF, Epo, interleukins. [Pg.203]

Third type of culture is stroma-non contact . In this system primitive progenitor cells are sustained when cells are co-cultured with irradiated allogeneic stroma but separated from it by the 0,4 micron membrane in transwell inserts (Costar, Cambridge, MA). These cultures are maintained by daily supplementation of stromal feeder conditioned media (Roller et al. 1998, Verfaillie, 2001) successfiilly expanded umbilical cord blood cells in a novel automated perfusion culture system. Development these approaches followed in the studies of investigators who incorporated the stromal components into the expansion culture. Recently published trials by McNiece et al. 2000 are more encouraging where cells were expanded in static culture for 10 days in Teflon bags (American Fluoroseal, USA). [Pg.205]

The first batch of cells consisted of AC 133+ cells cultivated in the diffusion chambers submerged on top of the feeder (feeder -AC 133 cells /Fl-C). The second batch consisted of human embryonic liver cell suspension directly cocultured with AC133+ cells at equal initial quantities (5x10 ) in the diffusion chambers submerged in the 6-well plates without additional feeder layer (FC-C). Third batch of experiments represented AC133+ cells cultured in the DC surrounded by FL condition media (condition media-AC133+ cells/CM-C). In the control group cells were cultivated in the same condition without any additions and without feeder layers. [Pg.206]

Our data suggests that hematopoiesis can be sustained for prolong cultivation periods in the presence of feeder layer cells or condition media supported culture models. Prolonged support of primitive hematopoietic cells (undifferentiated cells such as promyelocytes, myelocytes and metamyelocytes) and their clonogenic capacity and functional characteristics in feeder layer positive cultures, indicates that diffusible factors are sufficient and that direct cell-to-cell contacts may not be exclusively required for successful long term in K/fro hematopoiesis. [Pg.207]

Huang, R.R, Huang, R., Fan, Y., and Lin, Y, Simultaneous detection of multiple cytokines from conditioned media and patient s sera by an antibody-based protein array system. Anal. Biochem., 294, 55-56, 2001b. [Pg.235]

Primary cultures developed from pig and cow tissue are the best studied [29-32]. These models closely resemble the BBB, exhibiting many of the key biological properties. However isolation of blood-brain endothelial cells requires relatively complex cell isolation procedures which are labor-intensive and not ideal for screening purposes. Other cell types have been shown or proposed to induce barrier function, for example, astrocytes/pericytes. Significant improvements in barrier function was achieved in these primary culture models by including astrocyte conditioned media or co-culturing with astrocytes [33]. The complexity of primary cultures led to the use of epithelial cell lines not derived from the BBB (e.g., MDCK, MDCK-MDRl or LLC-PKl) [33]. [Pg.123]

NT419 Rennenberg, H. Glutathione in conditioned media of tobacco suspension cultures. Phytochemistry 1976 15 1433-1434. [Pg.361]

In 1985 Nerup and co-workers demonstrated that treatment of isolated rat and human islets with conditioned media derived from activated mononuclear cells resulted in the inhibition of glucose-stimulated insulin secretion (Mandrup-Poulsen et cd., 1985). The cytokine IL-1/3 was found to be the active component of the conditioned media (Bendtzen etal., 1986). Mandrup-Poulsen et al. (1986) further showed that continuous exposure of islets to IL-1 is toxic. [Pg.181]

Fig. 18.3. Raman spectral analysis of foetal osteoblast (FOB) differentiation. Unsupervised PCA of FOB cells cultured for 3 days in bioactive glass (BG) conditioned media (triangle) or control media (circle) (a). BG-treated cells formed a distinct cluster separate from control cells after 3 days culture. Least square (LS) analysis (which decomposes the cell spectra into the linear combination of Raman spectra obtained from the pure chemical constituents of the cell, e.g. nucleic acid, proteins, lipids, phospholipids and carbohydrates) of the relative RNA concentration of FOBs cultured for 1, 3 and 14 days in culture media (black) or BG condition media (grey), revealed a significantly reduced relative RNA concentration in FOBs culture in BG-conditioned media (b). FOBs cultured in BG-conditioned media appeared to accelerate FOB differentiation into mature adult osteoblast phenotypes (parallel gene and protein expression experiments confirmed this). Significant difference to control (p <0.05) [38]... Fig. 18.3. Raman spectral analysis of foetal osteoblast (FOB) differentiation. Unsupervised PCA of FOB cells cultured for 3 days in bioactive glass (BG) conditioned media (triangle) or control media (circle) (a). BG-treated cells formed a distinct cluster separate from control cells after 3 days culture. Least square (LS) analysis (which decomposes the cell spectra into the linear combination of Raman spectra obtained from the pure chemical constituents of the cell, e.g. nucleic acid, proteins, lipids, phospholipids and carbohydrates) of the relative RNA concentration of FOBs cultured for 1, 3 and 14 days in culture media (black) or BG condition media (grey), revealed a significantly reduced relative RNA concentration in FOBs culture in BG-conditioned media (b). FOBs cultured in BG-conditioned media appeared to accelerate FOB differentiation into mature adult osteoblast phenotypes (parallel gene and protein expression experiments confirmed this). Significant difference to control (p <0.05) [38]...
Kozma s review was a response to Clark (1983), who asserted that media do not influence learning under any conditions. .. Media are mere vehicles that deliver instruction but do not influence student achievement any more than the truck that delivers our groceries causes changes in our nutrition. According to Clark, the mode of presentation may affect the cost or extent of the distribution of education, but only the content of the vehicle can influence achievement (p. 445). Clark cited many reviews and studies that concluded that media do not affect learning and that many studies that compared media had been fruitless. [Pg.228]

Scale-up of cell cultures makes use of suspension cultures (erythropoietic cells or microcarriers) or, less often use of capillary beds (hollow fibre systems or glass bead columns), but these suffer from the same disadvantages seen with smaller scale cultures ( 3.4.4). In particular, nutrients are depleted as the medium flows through long columns or beds and high rates of flow coupled with recirculation are often employed. Nevertheless, Organon have used a hollow fibre dialysis system for production of monoclonal antibodies (Schonherr et al., 1985). Invitron s hollow fibre system has been used to produce cell conditioned media and the Cell-Pharm System (Jencons Ltd. Appendix 3) will produce up to 20 g cell secreted product per month. [Pg.56]

A 50 50 mixture of fresh growth medium with one of the above conditioned media is often recommended for growth of embryonic stem cell cultures. Otherwise the protein fraction can be concentrated for addition to serum-free medium. [Pg.93]


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See also in sourсe #XX -- [ Pg.151 ]




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