Fibroblast cultures

Moy LS, Howe K, Moy RL (1996) Glycolic acid modulation of collagen production in human skin fibroblast cultures in vitro. Dermatol Surg 22(5) 439-441... 

Watanabe N, Ando K, Yoshida S, et al. Gene expression profile analysis of rheumatoid synovial fibroblast cultures revealing the overexpression of genes responsible for tumor-like growth of rheumatoid synovium. Biochem Biophys Res Commun 2002 294(5) 1121-1129. 

Defects have been found in these mechanisms that cause various human diseases. For example, patients with the genetic disease xeroderma pigmentosum are especially sensitive to ultraviolet light and develop skin cancer. Skin fibroblasts cultured from these patients have been shown to be defective in DNA repair. 

Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126 663-676... 

The antiretrovirus properties of NO were shown in mice infected with Friend leukemia virus, a murine retrovirus. NO produced by NO-generating compounds or activated macrophages inhibits viral replication in fibroblast cultures, and is involved in defens against this murine retrovirus in vivo [134]. It was also reported that NO donors can inhibit HIV-1 replication in human monocytes through induction ofiNOS [135],... 

Arechabala, B., Coiffard, C., Rivalland, P., Coiffard, L. J. M. and de Roeck-Holtzhauer, Y. (1999). Comparison of cytotoxicity of various surfactants tested on normal human fibroblast cultures using the neutral red test, MTT assay and LDH, J. Appl. Toxicol., 19, 163-165. 

Several diaryl teUurides exhibit protection against TBH-induced cell death in lung fibroblast cultures. Besides, the same compounds prevent leucocyte-mediated cell damage in Caco-2 cells and protect rat kidney tissue against oxidative damage caused by anoxia and reoxygenation. 

Beside this dermatoxic activity pederin (147) has various biological activities (92). When administered in appropriate doses to partially hepatectomized rats, this compound stimulates development of hepatic tissues. The inhibitory effect at the cellular level has been found in chicken heart fibroblast cultures, and mice embryo, dog kidney, HeLa, and KB cell lines. In plants, root growth of Lupinus albus is inhibited and mitosis in Allium cepa blocked at the metaphasic stage. Also, pederin (147) inhibits protein synthesis and growth of yeast cells. In addition, the treatment of rat ascites sarcoma with purified extracts of P. fuscipes produces almost complete regression. 

Saito, K. Fibroblast cultures. Nippon Yakugaku Zasshi 1936 23 1-5. Harborne, J. B., and C. A. Willliams. 

Savage, J.R.K. and Woods, A.C. Differential effects of sulphur mustard on S-phase cells of primary fibroblast cultures from Syrian hamsters. Mutat. Res. 84 375-387, 1981. 

Nagai, N., Mori, K., Satoh, Y., Takahashi, N., Yunoki, S., Tajima, K., and Munekata, M. (2007). In vitro growth and differentiated activities of human periodontal ligament fibroblasts cultured on salmon collagen gel.. Biomed. Mater. Res. 82A, 395-402. 

Minimal essential medium (MEM) with Earl s salts and L-glutamine (Sigma M4655). Note complete MEM is used during fibroblast culturing, but the in vitro probe medium (IVPM see below) is used during the 72-h incubation described in the procedure. 

Fibroblast cultures for each patient and control are grown to confluency in T-75 culture flasks. The assay is then performed in triplicate using six-well plates. First, 1 ml of 0.25% trypsin-ethylenediaminetetraacetic acid (EDTA) solution (Sigma T4049) is added to each T-75 flask. After the cells have lifted from the flask, they are resuspended in 6 ml complete MEM. A 1-ml aliquot of the cell suspension is transferred to a 50-ml conical tube containing 14 ml of MEM complete medium (see above). Following further resuspension, 4 mL is plated into each of three wells in a six-well culture plate and placed into an incubator. Once the cells have attached to the seeded plates and are approximately 90-100% confluent (3-4 days), complete MEM is replaced with 1.5 ml of complete IVPM (see above). 

Table 3.2.7 Mass assignment and reference range for acylcarnitine species accumulating in fibroblast cultures following incubation with palmitate, liCa-valine, liC>>-isoleucine, and L-carnitine...

Fig. 3.2.8a-c Profiles of acylcarnitines as their methyl esters in cell culture medium (precursor of m/z 99 scan) following the in vitro probe assay in fibroblast cultures of a normal control (a) and a patient with the milder (b) and the more severe variant (c) of VLCAD deficiency. Note the more prominent elevation of dodecanoyl- (Ci2 m/z 358 peak 1) and myristoylcarnitine (Ci4 m/z 386 peak 2) compared to a relatively normal accumulation of palmitoylcarnitine ( , m/z 416 peak 3) in the milder VLCAD variant compared to the severe variant, where palmitoylcarnitine is markedly elevated. The asterisks represent the internal standards (from left to right) d3-acetyl-carnitine (C2 m/z 221), d9-isovalerylcarnitine (C5 m/z 269), d3-octanoylcarnitine (C8 m/z 305), d3-dodecanoylcarnitine (Ci2 m/z 361), and d3-palmitoylcarnitine ( m/z 419)... 

Diagnosis of HCS deficiency can be made by showing multiple carboxylase deficiency in skin fibroblasts cultured in low biotin medium but normal or clearly higher activities in high biotin medium [2, 30]. 

Cells Lymphoblasts or primary skin fibroblasts cultured for 4-7 days in lipoprotein-depleted medium, harvested by centrifugation and used fresh or after storage as pellets at - 18°C or lower. The minimum amount required for analysis is = 1.10 cells. 

Dichloromethane induced DNA single-strand breaks in mammalian cell cultures, but inconclusive or negative effects were reported for induction of gene mutations. It did not induce unscheduled DNA synthesis either in vivo in rodents or in human fibroblast cultures. It was genotoxic in fungi but not in Drosophila in the sex-linked recessive lethal assay. 

Physarum phospholipids (PHYLPA), isolated from myxoamoebae of Physarum polycephalum, is an inhibitor of eukaryotic DNA polymerase [71]. PHYLPA inhibits the proliferation of human fibroblasts cultured in a chemically defined medium. 

Fetal hamster fibroblast cultures (lethally irradiated feeder layer) ++ No... 

However, the concentration of HA in the ECM can vary widely. Even when the levels are decreased, as in areas of marked fibrosis, HA functions as an organizer of the ECM, as a scaffold about which other macromolecules of the ECM orient themselves. Diameters of collagen fibers can be modulated by levels of HA, the thinner more delicate fibers being favored in regions of high HA concentrations. In fibroblast cultures, the addition of exogenous HA to the medium decreases the diameter of the collagen fibers that accumulate (unpubl. observ.). 

Lever, J.E., Clingan, D., Jimenez de Asua, L. (1976). Prostaglandin F2alpha and insulin stimulate phosphate uptake and (Na+, K+) ATPase activity in resting mouse fibroblast cultures. Biochem. Biophys. Res. Commun. 71, 136-142. 

The biopsy material is used to obtain biochemical, morphological, and genetic information to establish a diagnosis. Mitochondria isolated from the fresh muscle biopsy or from cells (fibroblasts) cultured from the skin biopsy can be used to measure respiratory chain activity by polarography. This procedure measures 02 consumption as different respiratory chain substrates are added to the mitochondrial suspension. 

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