Skin fibroblasts, incorporation

Incorporation of tritium (cpm/mg of protein) into individual moieties of glycosphingolipids of normal human skin fibroblasts in the presence and absence of galactose oxidase... 

HGPRT activity in L.W. was less than 0.01 nmol/mg HbA in lysed red cells, less than 0.9 nrool/mg proteinA in skin fibroblasts and less than 0 2% of normal in intact erythrocytes. Table 2. Incorporation of labelled hypoxanthine into purine nucleotides by intact fibroblasts was approximately 6% of control which is low on the criteria of Page et al. These results confirm severe HGPRT deficiency. Heterozygote levels were found in the Mother s fibroblasts... 

Hyaluronic acid can be re-incorporated into some cells that synthesize it. This adsorptive pinocytosis occurs with mesenchymal cells, but not with either skin fibroblasts or arterial smooth-muscle cells. cAMP appeared to stimulate the synthesis of hyaluronic acid by human fibroblasts and confluent rat fibroblasts. ... 

Slide 4 (Fig.4) Here HG-PRT negative human skin fibroblasts were grown on coverslips. Normal erythrocytes preincubated with %-hypoxanthine were added to the culture medium and were allowed to settle down on top of the fibroblasts The erythrocytes were then poured off, and the few remaining red cells lysed by hypotonic shock Autoradiography reveals incorporation of radioactive label in the HG-PRT negative fibroblasts ... 

Chalmers and Spellacy (1980) studied cultured skin fibroblasts from this child and were unable to demonstrate deficient activity of 3-methylcrotonyl-CoA carboxylase, or of propionyl-CoA carboxylase measured as a control, in disrupted confluent fibroblasts. However, using a newly developed assay that measures incorporation of [l- C]isovaleric acid into protein, with incorporation of [l- C]acetyl-CoA as control, in intact subconfluent dividing fibroblasts grown in monolayer (Chalmers and Spellacy, 1979), which effectively screens for disorders in the metabolism of L-leucine between isovaleryl-CoA and acetyl-CoA, these workers were able to demonstrate a grossly reduced incorporation of isovalerate into protein (21 and 23 pmol of... 

The incorporation of cellular growth-enhancing substances or additional cell types, besides keratinocytes and fibroblasts, is also achieved to obtain constructs with improved functions and higher resemblance to native skin. 

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