Chemistry: molecular biology and microbiology – Spore forming or isolating process
Patent
1981-02-18
1985-08-06
Rosen, Sam
Chemistry: molecular biology and microbiology
Spore forming or isolating process
435 1, 435241, 128334R, C12N 500
Patent
active
045336350
DESCRIPTION:
BRIEF SUMMARY
This invention relates to processes and products applicable to the human skin and more particularly to the adult human epidermis. Its object is notably a process for stimulating the growth of human epidermal cells, particularly those of adults, as well as cosmetic, pharmaceutical and diagnostic products using the said process.
The skin is the principal tissue exposed to the mutagenic effects of chemical and physical environmental agents. This is particularly true of radiations of every sort. The skin is composed of tissues of mesenchymal origin (the derma and the blood vessels) and tissues of ectodermic origin (the epidermis). Now, although it is known to extract mesenchymal cells (fibroblasts) from the dermis to culture them and to study the mutagenic and carcinogenic effects of chemical and physical environmental factors, it is difficult to extract them from the epidermis. One of these difficulties had not yet been overcome. This is the application of processes of cytogenetic examination to cultured adult epidermal cells, which involves on the one hand the preparation and analysis of chromosomes, and on the other, the notation of sister chromatid exchanges (SCE). In point of fact, although a few rare publications mention succinct chromosomic investigations, there is no exhaustive chromosomic investigation, particularly in so far as heriditary human skin diseases are concerned and, to the best of the applicant's knowledge, no bibliographical references exist with respect to SCE.
A series of recent investigations suggests that the increase of SCEs is a sensitive and accurate indication of the mutagenic effect of a chemical product or of a radiation. The application of SCE-revealing techniques to adult human epidermal cells is, therefore, unquestionably important.
The object of the invention is, notably, a process applicable to the preparation and the analysis of chromosomes and to the preparation and the notation of SCEs. According to the invention, it is possible to induce mitotic multiplication of these cells on a normally hostile glass substrate.
The invention also makes it possible to provide a solution to all the problems connected with the growth and culture of epidermal cells, particularly in man and especially in adults. Thus, apart from the abovementioned cytogenetic application, the invention can also be used in the treatment of burns, in the fields of cosmetics and of cutaneous pharmacology and for long-term culture of epidermal cells.
The present invention calls upon previous work reported by ARRUTI C. and COURTOIS Y. Exptl. Cell. Res. 117, 283-292 (1978). These authors established that a retinal extract, referred to by the abbreviation RE, was a growth-promoting factor for cultured epithelial lens cells. The skilled artisan can, if needs be, refer to this article to obtain the necessary information on this retinal extract RE, and the properties which have been observed. The authors used a total retinal extract obtained by extraction with an aqueous salt solution, with a pH 7.2 phosphate buffer such as the type sold by FLOW Laboratories Ltd. The bovine retinas are placed in intimate contact with such a solution. After a certain number of physical centrifugation and filtration techniques the retinal extract RE is isolated. However, the observations reported in this article are limited to the growth of epithelial lens cells.
Certain documents relating to the prior technique describe work on extracts obtained from animal ocular tissue.
French Pat. No. 71.13.756 (publication 2.134.088) describes a process for obtaining a total bovine eyeball extract. The series of steps involves the action of merthiolate, of an alcaloid and of "Celite".
The alcoholic extract so obtained is applied in the field of ophthalmology. It is not, therefore, an aqueous saline solution, and its use for the growth of epidermal cells is not mentioned.
The article by B. S. Kasavina et al. (Columbus, Ohio, USA) and SU Pat. No. 130.159 of the July 15, 1960 cited in Chemical Abstracts Vol. 55, No. 5 (Mar. 6, 1961) No. 4892c descri
REFERENCES:
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Arruti Cristina
Barritault Denis
Courtois Yves
Guedon born Saglier Isabelle
Prunieras Michel
Institut National de la Sante et de la Recherche Medicale
Rosen Sam
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