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Biomedical subjects

L I Huschtscha

Publications and source records attributed to L I Huschtscha.

6 recordsLinked to original sources
PubMed1989

A rapid micro method for counting cells "in situ" using a fluorogenic alkaline phosphatase enzyme assay.

A new method has been developed to count cells "in situ", based on a fluorogenic enzyme assay that measures the activity of alkaline phosphatase. Increasing cell number was shown to correlate closely with alkaline phosphatase activity and this relationship did not change with time in culture. The alkaline phosphatase assay (ALP assay) was able to estimate relative cell numbers over a range from about 10(4) to 5 X 10(5) for many cell types, including Hep-2, a derivative of HeLa, several human colorectal cell lines SW1222, SW837, LS174T and HT29, a normal human diploid cell strain MRC5 and a rodent line NIH-3T3. The ALP assay is rapid and efficient, making it a useful method for studying growth assays.

Alkaline Phosphatase↗
PubMed1986

The susceptibility of Werner's syndrome and other human skin fibroblasts to SV40-induced transformation and immortalization.

In attempts to transform and immortalize human cell cultures, skin fibroblasts from normal donors of different ages, from patients with the premature ageing diseases Werner's syndrome (WS) and progeria (PR), and from donors with the cancer-prone diseases ataxia telangiectasia (AT), Bloom's syndrome (BS) and Fanconi's anaemia (FA), were infected with SV40 virus and their growth monitored thereafter. Lesch-Nyhan (LN) fibroblasts were also infected. SV40-infected cultures from two normal and from WS, AT and LN donors attained a spectrum of transformed properties, high mitotic activity at confluence, presence of T-antigen, anchorage independence and altered morphology. Most of these pretransformed cultures died in the crisis period. However, two cultures from the WS and LN patients survived the crisis period and have now been grown to more than 200 passages. For the LN culture the crisis period was at least 200 days. Both permanent lines retain the properties of pretransformed cells, but differ in their modal chromosome number and ability to grow in methionine-free medium. It can be concluded from these experiments that transformation by SV40 to permanent lines is a rare event in human skin fibroblasts, even when these cells were taken from patients predisposed to form cancers.

Age Factors↗
PubMed1977

Testing the commitment theory of cellular aging.

The commitment theory may explain both the finite lifespan of diploid fibroblasts and the apparent immortality of transformed lines. Potentially immortal cells are assumed on division to generate with some fixed probability cells committed to senesce after a specific number of divisions. During the period between commitment and senescence, cells are assumed to maintain normal growth so that the uncommitted cells are diluted by committed ones and may ultimately be lost in subculturing. A number of predictions of this model are described and experiments strongly supporting the theory are reported. We conclude that the limited growth of diploid fibroblasts is, in effect, an artifact of normal culturing procedures.

Cell Cycle↗
PubMed1977

Age changes of the pattern of F1 histone subfractions in rat liver and spleen chromatin.

The polyacrylamide gel electrophoresis of total histone extracted from young (3 months) and old (26-27 months) rat tissues does not show age-related differences in the pattern of the main five histones. However, when the lysine-rich F1 histone was extracted from chromatin separately by perchloric acid and purified by Biogel P-60 column chromatography from chromatin of rat liver and spleen, the polyacrylamide gel electrophoresis did show obvious age changes in the distribution of F1, F1o, and F1(00) subfractions. In liver chromatin the increase of rat-specific methionine-containing subfraction (F1met.) was also found.

Aging↗