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Biomedical subjects

M Eisinger

Publications and source records attributed to M Eisinger.

49 records · Page 3Linked to original sources
PubMed1982

Selective proliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin.

Cultures consisting almost entirely of human melanocytes were obtained from epidermal single-cell suspensions by using phorbol 12-myristate 13-acetate (10 ng/ml) in the culture medium. At this concentration, phorbol ester is toxic to human keratinocytes but not to melanocytes. When the seeding density was optimal (0.8-2 x 10(4)/cm2) and the medium contained both phorbol ester and cholera toxin, melanocytes proliferated extensively. Under these conditions, human melanocytes could be passaged serially in vitro and grown in quantity. This cell culture system can thus be used to answer basic questions related to pigment cell biology and may serve as a control for studies of malignant melanocytes.

Cell Division↗
PubMed1982

Immunoelectron microscopic identification of Langerhans cells using a new antigenic marker.

The specificity of a monoclonal antibody (OKT6) for epidermal Langerhans cells was examined by immunoelectron microscopy. Peroxidase-labeled OKT6 bound to 1-5% of suspended human epidermal cells, as determined by light microscopy. Electron microscopic examination of peroxidase-labeled cells revealed that all Birbeck granule-containing Langerhans cells bound OKT6. In addition, a small population of indeterminate cells, lacking the Birbeck granule, was also labeled with OKT6. The ultrastructural studies confirm the specificity of OKT6 for Langerhans cells and suggest that the indeterminate cell represents a related cell population.

Antibodies, Monoclonal↗
PubMed1981

Pemphigus, pemphigoid, and epidermal upper-cytoplasmic antigens: changes in expression in cultured human keratinocytes.

In an approach of epidermal differentiation, the expression of pemphigus, bullous pemphigoid, and upper-cytoplasmic epidermal antigens was studied in human keratinocytes in culture. The cells were cultured without feeder cells, dermal tissue, or collagen at an acid pH (5.6--5.8) similar to that of the surface of the skin in vivo. Cell suspensions from fresh trypsinized skin and primary, secondary, and tertiary cultures were tested by indirect immunofluorescence for the presence of each antigen using human sera from patients with pemphigus, bullous pemphigoid, and human sera with antibodies against upper-cytoplasmic antigens. Normal sera and cultured human normal fibroblasts and melanoma cells were used as controls. Pemphigus and pemphigoid antigens were found to be expressed, and synthesized by keratinocytes in vitro. The expression to upper-cytoplasmic antigens decreased with time in culture, and they were absent in secondary or tertiary cultures, while expressed by 45--65% of cells prepared from fresh skin. Both upper-cytoplasmic and pemphigoid antigens can be used to type subpopulations of human epidermal cells; however, these findings suggest that epidermal differentiation in vitro differs from that which occurs in vivo.

Antigens↗
PubMed1981

Metabolism of benzo[a]pyrene and DNA adduct formation in cultured human epidermal keratinocytes.

Cultured human epidermal cells which require no feeder layer were used to study metabolism of benzo[a]pyrene (BP) and DNA adduct formation. The cultures were prepared from a single cell suspension and maintained at a pH of 5.9--6.2. At 2 microM BP some cell toxicity was observed, and substantial cell death occurred at 4 microM BP. The metabolism and DNA binding of BP were followed from 6 to 48 h of incubation. High pressure liquid chromatography (h.p.l.c.) revealed that BP was metabolized into 9,10-diol, 7,8-diol, quinones, phenols and tetraols of BP. The DNA binding levels increased linearly up to 28 h of incubation. At 18 h, the level of DNA binding at 0.4 microM BP was 1.5 x 10(-6) mol BP/mol DNA and increased to 6.0 x 10(-6) mol BP/mol DNA at a dose of 4 microM BP. Analysis of the DNA adducts by h.p.l.c. indicates that the 2'-deoxy-N2-(7,8,9,10-tetrahydro-7 beta-,8 alpha, 9 alpha-trihydroxybenzo[a]pyrene-10-yl) guanosine was the predominant adduct formed in cells exposed to BP. The prevalence of the minor DNA adducts varies as a function of the source of the primary skin cells. These results confirm that human cells with no feeder layer metabolize BP and the resultant DNA damage is similar to that found in other mammalian systems.

Benzo(a)pyrene↗
PubMed1981

Decreased expression of epidermal cytoplasmic antigens in cultured human keratinocytes.

The expression of upper cytoplasmic (U-CYT) antigens which are expressed only in the superficial layers of the epidermis and are markers of epidermal cell differentiation in vivo and of basement zone (BMZ) antigens reacting with bullous pemphigoid serum was studied in keratinocytes in tissue culture. The cells were cultured at an acid pH (5.6-5.8) similar to that of skin and without feeder cells, dermal tissue, or collagen. It was found that the expression of U-CYT antigens decreased markedly in culture. These antigens were expressed in 45-65% of epidermal cells prepared from fresh skin, but in only 5-10% of cells which had been grown in primary culture over 1 mo, and in no cells in secondary or tertiary culture. By contrast, BMZ antigens continued to be expressed in culture. These antigens were expressed by 20-35% of epidermal cells prepared from fresh tissue and by 15-35% of keratinocytes in primary, secondary or tertiary culture. These findings indicate that U-CYT and BMZ antigens can be used to type subpopulations of human keratinocytes in suspension, and suggest that the differentiation of these cells in vitro differs from that which occurs in vivo.

Antigens↗
PubMed1980

Wound coverage by a sheet of epidermal cells grown in vitro from dispersed single cell preparations.

Epidermal cells have been grown into multilayered sheets resembling full-thickness human epidermis using a new tissue culture technique. To evaluate their viability in an in vivo situation, such cells were transplanted under the kidney capsule and into a subcutaneous position in the nude mouse. In both locations cells survived and limited growth was observed. Maturation of the epidermal cells resulted in the appearance of keratohyalin granules and cell desquamation. To assess the potential use of tissue cultured cells for wound treatment in man, the same method was used to grow dog cells in vitro. Sheets of multilayered dog epidermal cells grown in vitro from a single cell suspension were used to cover fresh or granulating wounds. The epidermal cells transplanted in this way remained viable and wound coverage was achieved within 1 week. No clinical or histological signs of rejection of allograft tissue were observed during a 6-week period of observation.

Animals↗
PubMed1979

Human epidermal cell cultures: growth and differentiation in the absence of differentiation in the absence of dermal components or medium supplements.

Human epidermal cells grew and differentiated in vitro, provided that the pH of the culture medium was at 5.6-5.8, the seeding density was optimal (approximately 2.5 x 10(5) cells per cm2), and the incubation temperature was maintained at 35-37 degrees C. Under these conditions, epidermal cells from many different skin locations grew to confluency within 15-20 days and formed multi-layered sheets whose differentiated structure resembled that of the full depth of skin epidermis. Cell proliferation and differentiation did not require a feeder layer, a collagen substrate, a high concentration of fetal bovine serum, or added hormones. The sheets of differentiated epidermal cells could be dissociated from the plastic surfaces of the tissue culture flasks. The use of such cultured cells for wound dressing is proposed.

Cell Differentiation↗
PubMed1978

A quantitative micro-complement fixation method in studies of human wart viruses.

This report offers a description of a quantitative micro-complement fixation method (Cikes, 1975) for detecting human wart virus antigens and their specific antibodies, and proof of the specificity of the reactions being detected. The increased sensitivity demonstrated by chromium-release measurement is compared to the results of visual interpretation of complement fixation.

Antibodies, Viral↗
PubMed1977

Identification of an immunologically distinct papillomavirus from lesions of epidermodysplasia verruciformis.

Virions isolated from warts of 2 siblings with epidermodysplasia verruciformis (EV), a rare disease characterized by the lifelong growth of warty skin tumors containing papova-like virions, were compared to isolates of human papillomavirus (HPV) from 3 pools of plantar and common hand warts. The length of relaxed, circular (form II) molecules of EV virion DNA approximated the length of HPV DNA but was 3.3% longer. Antisera prepared in rabbits against the 3 HPV pools coated and aggregated HPV in immune electron microscopy (IEM) tests but did not react with EV virions. These antisera reacted at high titers in complement fixation (CF) tests with HPV and reacted only weakly in CF tests with EV virions. Rabbit antisera to EV virions coated and aggregated EV virions but reacted only weakly or not at all with HPV virions in IEM tests. These sera reacted in CF with EV virions only. The data indicated that virions from the EV patients represent an immunologically distinct papillomavirus.

Antigens, Viral↗
PubMed1976

Cell-mediated immunity (CMI) to human wart virus and wart-associated tissue antigens.

Lymphocyte transformation (LT) and leucocyte migration inhibition in agarose were used to demonstrate cell-mediated immune response to purified human wart virus and wart tissue extract in various subjects with warts and those without past history of warts. Most individuals bearing warts for less than 1 year duration showed positive cell-mediated responses to both the virus and tissue extract whereas very few of those who had warts for longer duration responded to either antigenic preparation. The difference was statistically significant. Subjects who had warts in the past also showed positive responses but these tended to decrease in degree with time. Surprisingly a group of subjects who never had warts before also responded to stimulation with the virus, but not to the extract. The positive response to stimulation with wart tissue extract reflects the presence of wart associated antigens other than the virus. Cell-mediated immunity against the wart virus and wart-associated antigens is probably important in preventing the persistence or even establishment of disease but this protective immunity is short-lived. The lack of quantitative correlation between LT and leucocyte migration inhibition demonstrable in this study suggests that these two are separate events in in vitro lymphocyte stimulation with antigens.

Adult↗
PubMedDate not supplied

In vitro fibroblast seeding of prosthetic anterior cruciate ligaments. A preliminary study.

To evaluate the effect of in vitro seeding of fibroblasts on the connective tissue encapsulation of implanted ligament prostheses, canine skin fibroblasts were grown in tissue culture and seeded onto knitted Dacron prostheses. When the cells on the prostheses reached contact growth inhibition, as determined by growth curves, the prostheses were implanted into the dogs' knees as ACL replacements. Gross and histologic evaluation at 4 and 8 weeks revealed that the seeded prostheses consistently showed more uniform and abundant encapsulation with connective tissue than did the control (unseeded) prostheses. The giant cell response observed in the tissue surrounding the unseeded prostheses was not noted in the seeded prostheses. This may be because seeding a prosthesis with fibroblasts prior to implantation "walls off" the prosthesis from the environment of the joint, and therefore the prosthesis may not elicit as great a foreign body response as does an unseeded prosthesis. The results of this preliminary study suggest that the in vitro seeding of a ligament prosthesis with fibroblasts accelerates the connective tissue encapsulation of the implanted prosthesis.

Animals↗