DNA flow cytometry on human epidermis. The effect of serial biopsy sampling at various times

Frentz, Gerda; Møller, Ulla; Keiding, Niels

doi:10.1111/j.1365-2133.1982.tb00283.x

Cited by 9 publications

(2 citation statements)

References 14 publications

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“…Few publications that make reference to the study of normal skin by flow cytometry and are related to methods for evaluating the kinetics of epidermal cells. The location and local injure can alter the previous results of the study, but gender, age, time of day, length of storage in liquid nitrogen appear to be less important [10,[12][13][14]. The high risk basal cell carcinomas (n = 21), showed significantly enhanced the population tumor cells in phases of synthesis (17.8%), followed by G0/G1 phases (14.5%), apoptosis (9.1%) and increased cells in proliferation G2/M (7.0%).…”

Section: Discussionmentioning

confidence: 94%

Prognostic factors correlation between the cell cycle phases and apoptosis in basal cell carcinoma

Festa-Neto¹,

Maria

2011

OJIM

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Basal cell carcinoma (BCC) is the most common skin malignancy, are found in various forms depending on their clinical and biological behavior. The objective of study was analyzed the phases of the cell cycle and correlations between BCC of low and high risk of recurrence and correlation prognostic factors. The quantity of content DNA in tissues of normal skin, showed small amount of cells in apoptosis and mostly in phase quiescent and rare aneuploidy cells. In BCC, apoptosis was higher in the BCC at high risk than low risk, probably due to their high rates of cell proliferation, and present of aneuploidy cells, when compared to the average percentage of aneuploidy. The DNA content from cells of normal skin shows that the majority is in the quiescent phase; compatible with tissues that are is refreshing. There is presence of apoptosis in the epidermis by probable normal process of differentiation. The aneuploidy in BCC showed a direct correlation with the degree of tumor aggressiveness

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Section: Discussionmentioning

confidence: 94%

Prognostic factors correlation between the cell cycle phases and apoptosis in basal cell carcinoma

Festa-Neto¹,

Maria

2011

OJIM

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“…For the thirteen patients who provided biopsies from unexposed, exposed, and neoplastic skin, a two-way analysis of variance was carried out on the logarithmic values to evaluate whether the differences were due to individual factors or to differences depending on the category of skin studied. For evaluation ofthe correlation between the uninvolved skin and the tumour type, pre-existing conditions and carcinogen exposure, analyses of variance were carried out using various models depending on the importance ofthe different variables (Frentz et al, 1982). The 5"o probability level was used.…”

Section: Flow Cytometry and Statistical Analysismentioning

confidence: 99%

DNA flow cytometry of the epidermis of patients with multiple epidermal carcinomas

Frentz

Møller²

1984

Br J Dermatol

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We used DNA flow cytometry to determine whether the uninvolved skin of twenty-four patients with multiple epidermal cancers differed from normal skin with regard to its epidermal cell kinetics. We also studied the relationship between the cell kinetics and the type of cancer, previous exposure to ionizing or UV radiation, previous arsenic intake, skin type and coexisting psoriasis. Both exposed and unexposed skin was studied. In thirteen patients the S and G2M fractions of a monoclonal tumour were compared with the values in unexposed and exposed skin. The S and G2M fractions increased progressively from unexposed skin to exposed skin to tumour tissue. Patients who had developed both squamous and basal cell carcinoma differed from those who had developed only basal cell carcinoma by having a significantly higher G2M fraction in the exposed skin, and significantly higher S and G2M fractions in the unexposed skin. Patients exposed to ionizing radiation tended to show higher G2M fractions in exposed skin compared with the remaining individuals (P = 0.06). The presence of psoriasis, exposure to UVR or sunlight and previous arsenic ingestion did not appear to affect the results.

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Modification of a trypsin‐detergent method for DNA flow cytometry of human epidermis

et al. 1989

Cytometry

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An existing technique (Vindelov et al. Cytometry 3:323, 1983) has been modified for DNA flow cytometry of human epidermis obtained from 2 to 3 mm punch biopsies. By varying the length of time of digestion of the epidermal disc by trypsin from 5 to 70 Key terms: Trypsin digestion, biopsy, keratimin a controlled release of keratinocytes occurred beginning with the stratum basale and proceeding toward, but not including, the superficial layer of the epidermis, the stratum corneum. nocyte, isolated nucleiAn important prerequisite for performing flow cytometry (FCM) is the acquisition of a single cell or nuclear suspension. For epidermis, the production of single cell suspensions permits the study of a variety of cell types such as Langerhans cells, keratinocytes, keratinocyte subpopulations, and basal cells (2-11). Because of our interest in the possibility of using cell cycle kinetic data obtained from human epidermis as indicators of the best circadian time for maximal host tolerance to chemotherapy, i.e., chronochemotherapy (11, we modified a n existing technique (12) for use in DNA FCM of human epidermis.Dispersion of epidermal cells or their nuclei into suspension has been accomplished by several different methods including trypsin, dithiotheritol (DTT), EDTA, ultrasonication, or ultrasonication plus DTT (2). The trypsin detergent method described by Vindelov et al. (12) has the advantages of (a) frozen storage of the specimen, (b) production of clean nuclei after digestion of cytoplasmic proteins with trypsin, and (c) quantitative staining of DNA with propidium iodide.This report describes a modification of an existing technique (12) for the production of isolated nuclei of keratinocytes obtained from 2-3 mm punch biopsies of adult human epidermis. Based on the duration of the incubation time in trypsin, a controlled release of keratinocyte populations can be achieved. Trypsin digestion dissolved epidermal cell cohesion beginning in the basal cell layer with progression toward the superficial-most layer after longer incubation. MATERIALS AND METHODS BiopsiesThe skin biopsies used in this study were obtained from the scalp during hair transplantation surgery. Epidermal-Dermal Separation and Freezing of Isolated EpidermisTo facilitate the removal of epidermis from dermis, skin biopsies were first individually placed in 0.5% cold acetic acid for 48 h. With the aid of a dissecting microscope, the epidermis was separated from the dermis using watchmaker's forceps. The disc of epidermis was then placed in fresh DMSO solution and frozen rapidly to -70" C. Intact biopsies as well as the peeled dermis and epidermis were stained with hematoxylin and eosin (H & E) and examined by routine light microscopy. Trypsinization and StainingThe trypsin-detergent method of Vindelov (12) was used with the modification that digestion times in trypsin were varied from 5 to 70 min. In addition to varying the digestion time, other variables which were studied included mechanical vortexing at 1 min, 3 min and 5 min intervals during th...

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DNA flow cytometry on human epidermis. The effect of serial biopsy sampling at various times

Cited by 9 publications

References 14 publications

Prognostic factors correlation between the cell cycle phases and apoptosis in basal cell carcinoma

Prognostic factors correlation between the cell cycle phases and apoptosis in basal cell carcinoma

DNA flow cytometry of the epidermis of patients with multiple epidermal carcinomas

Modification of a trypsin‐detergent method for DNA flow cytometry of human epidermis

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