OBJECTIVETo compare the attachment and growth characteristics between human junctional epithelium (JE) and oral epithelium cells.

METHODSThe healthy JE biopsies were derived from the human teeth extracted due to impaction or orthodontic purpose. Enzyme digestion was used to isolate JE cells, which were then cultured in DKGM. The co-culture model of JE cell-tooth slice was built up by adding 3 decalcification cementum slices (5 mm x 3 mm x 1 mm) into sterilized plate containing 1 ml of JE cells (5 x 10(8)/L), 21 slices all together,and incubated in an atmosphere containing 5% CO2 at 37 degrees C for 1-14 days. The attachment structure was observed under transmission electron microscope, and the OE cells was used as control.

RESULTSThe human JE cells were polymorphous in shape and CK19 positive, while OE cells were consisted of equal and closely packed epithelial-like cells in a paving stone arrangement, and CK19 was only strained in a few cells. There were a few cells in JE-slice when co-cultured for 1-3 days, and electron dense plaques on the JE cell surface of the attached slice were observed at 9 days, and 2-3 layer of JE cells and hemidesmosome-like structure formed within 11-14 days. There were more OE cells within 1-3 days, electron dense plaques appeared at 7 days, and stratified epithelium and hemidesmosome-like structure formed in OE-slice at 9 days.

CONCLUSIONSThe cultured JE cells were immature and lower differentiated epithelial cells which were different from OE cells. Under the same condition the growth and attachment of JE cells on the cementum slice surface were slower than that of OE cells. Their attachment strength needs further study.

Cell Count ; Cell Growth Processes ; Cells, Cultured ; Epithelial Attachment ; cytology ; Epithelial Cells ; cytology ; Gingiva ; cytology ; Humans

Neuroendocrine cells are abundant in all the body tissues and organs as well as the nervous system, either the central or the peripheral nervous system. In the normal prostate tissue, there are a few neuroendocrine cells, too, in addition to basal and epithelial cells. Prostatic neuroendocrine cells play the function of regulating the development, secretion and differentiation of the prostate. Recent studies show that prostatic neuroendocrine cells may be involved in the pathogenesis of chronic prostatitis through their activity and secreted products. This article presents an overview on the origin, distribution, morphology, structure, secretion and functions of prostatic neuroendocrine cells and their association with chronic prostatitis.
Chronic Disease ; Epithelial Cells ; cytology ; Humans ; Male ; Neuroendocrine Cells ; cytology ; Prostate ; cytology ; Prostatitis ; pathology

The aim of this study was to isolate, cultivate and phenotypically characterize two types of human amnio-tic membrane (HAM)-derived cells, and to analyze their differentiation potential in vitro. Human amnion epithelial cells (hAEC) were derived from the embryonic ectoderm, while human amnion mesenchymal cells (hAMC) were derived from the embryonic mesoderm. The cells were characterized by flow cytometry and immunofluorescence, then immunofluorescence also was performed for the analysis of multipotentiality in differentiation. The results indicated that immunophenotypic characterization of both cell types demonstrated positive for HLA-A, B, C and mesenchymal stem cell markers (CD29, CD73, CD44, CD59, CD90, CD105, CD166), but did not express the hematopoietic markers (CD31, CD34, CD45, HLA-DR) and showed the weak expression of costimulatory molecules (CD40, CD40L, CD80, CD86). Phenotypes of both cell populations were maintained from passages 3 to 7. The immunofluorescence indicated that hAEC expressed cytokeratin 19, but did not express vimentin. On the contrary, hAMC expressed vimentin but did not express cytokeratin 19. The assessment of multilineage potential demonstrated that hAMC showed greater cardiomyocytes potential, while hAEC showed greater neural potential. It is concluded that hAEC and hAMC can be successfully isolated from the HAM. Both cell populations possess similar immunophenotype. However, they differ in cell yield and multipotential for differentiation into the major lineages, hAEC possess a much greater ectodermal differentiation capacity, while hAMC possess a much greater mesodermal differentiation capacity. This conclusion will be important for use of these cells in cell therapy.
Amnion ; cytology ; Cell Differentiation ; physiology ; Cell Lineage ; Epithelial Cells ; cytology ; Humans ; Immunophenotyping ; Stromal Cells ; cytology

OBJECTIVETo explore the feasibility of the transdiferentiation of human breast adipose-derived stem cells (hbASCs) into mammary epithelial-like cells after co-culturing in Transwell in vitro.

METHODSThe third passage hbASC and the HBL-100 cell line were co-cultured in a Transwell culture system for 15 days. The hbASCs were observed and identified by inverted phase contrast microscope and transmission electron microscopy, and immunocytochemistry staining in the induced and control groups.

RESULTSBoth the third passage hbASCs and the HBL-100 cell line cells could adhere and grow rapidly after co-culture in the Transwell system. After co-culture for 15 days, the morphology of some induced hbASCs changed into epithelial-like cells. Some induced hbASCs showed positive expression of CK18, CK19 by immunocytochemistry staining, and typical epithelium cells with microvilli, desmosomes and tonofilaments observed under TEM. The positive rate of CK18 and CK19 was (24.4 +/- 12.0)% and (21.6 +/- 16.4)% in experimental group, and (1.8 +/- 1.7)% and (1.1 +/- 0.6)% in control group.

CONCLUSIONThe data suggests that hbASCs may have the potential to transdifferentiate into human mammary epithelial-like cells after co-culturing in Transwell in vitro.

Adipose Tissue ; cytology ; Breast ; cytology ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Epithelial Cells ; cytology ; Female ; Humans ; Stem Cells ; cytology

No abstract available.
Calcium Signaling/physiology* ; Cell Polarity/physiology* ; Epithelial Cells/physiology* ; Epithelial Cells/cytology*

It is commonly accepted that airway hyperresponsiveness (AHR) is a chronic airway inflammation although the exact mechanism of its pathogenesis is still unclear. In the past ten years, an epithelial defect hypothesis has gradually gained supports from the main stream. Airway epithelium is no longer considered only as a simple mechanic barrier but an active interface between the inner and outer environment. Bronchial epithelial cells play a critical role in maintenance of homeostasis in the airway local microenvironment through a wide range of physiologic functions including anti-oxidation, exocrine/endocrine secretions, mucus production and antigen presentation under health and stressed/inflamed/injured conditions. It is reasonably hypothesized that disruption of these functional processes or defects in airway epithelium integrity may be the initial steps leading to airway hyperresponsiveness such as in asthma and chronic obstructive pulmonary disease.
Animals ; Bronchi ; cytology ; Bronchial Hyperreactivity ; physiopathology ; Epithelial Cells ; pathology ; Humans

BACKGROUNDCervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS). However, it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor. Therefore, the culture of these cells is extremely difficult. In this study, we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro.

METHODSNormal cervical epithelial tissue pieces were obtained and digested with type I collagenase to dissociate the cells and a single cell suspension produced. The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type I from rat tail, with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS. After attachment, the medium were replaced with K-SFM without FBS. The expression of basal keratins of the ectocervical epithelium, K5, K14 and K19 were assayed by immunofluorescence with monoclonal antibodies to identify the cell purity.

RESULTSOur results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5% FBS than in K-SFM alone, as well as to tissue culture plastic coated with collagen type I than plastic alone. The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type I from rat tail was the best method for culture of normal cervical epithelial cells. K5, K14 and K19 were assayed and keratinocyte purity was nearly 100%.

CONCLUSIONA novel, simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

Cell Culture Techniques ; methods ; Cervix Uteri ; cytology ; Epithelial Cells ; cytology ; Female ; Humans ; Keratinocytes ; cytology

Animals ; Apoptosis ; Cataract ; etiology ; Cattle ; Cell Proliferation ; Cells, Cultured ; Epithelial Cells ; cytology ; Lens Capsule, Crystalline ; cytology

OBJECTIVETo investigate the primary culture of mammary epithelial cells derived from human breast hypertrophy.

METHODSMammary epithelial cells of human breast hypertrophy were isolated and cultured by the collagenase digestion method. The morphology observation and identification of the cultured cells were performed by inverted microscope observation, HE staining and cytokeratin immunohistochemical staining.

RESULTSMost of the cultured cells were pebbles-like or polygon under the inverted microscopy. Some had irregular form. They had typical island-like appearance during multiplication with close connection between the cells. The HE staining results showed the cytoplasm was stained pink or lilac, the nucleus was stained bluish violet and was round or oval in shape, with clearly visible chromosomes in dark blue. The cytokeratin immunohistochemical staining demonstrated the tissue-specific expression of cytokeratin 18 in epithelial cells by the cytoplasm stained claybank.

CONCLUSIONHigh purity of primary mammary epithelial cells derived from human breast hypertrophy can be obtained by the collagenase digestion method and conditioned medium.

Breast ; cytology ; pathology ; Cells, Cultured ; Epithelial Cells ; cytology ; Female ; Humans ; Hypertrophy ; pathology ; Primary Cell Culture ; methods

OBJECTIVETo study the human nasal ciliated epithelial cells at an air-liquid surface (ALI) so as to establish a reliable cell culture model for nasal mucociliary transport study.

METHODSThe human nasal ciliated epithelial cells were cultured by low-temperature enzymatic digestion method at an air-liquid surface, the cell growth behavior was observed under the inverted phase-contrast microscope, the proliferation, confluence and differentiation of cultured cells were examined by scanning electron microscope and immunocytochemistry, the basal and stimulated ciliary beat frequency (CBF) of cultured epithelial cells were measured by using high-speed digital microscopic imaging system. Prism 4.0 software was used to analyze the data.

RESULTS(1) Under microscope, the cells on transwell membrane adhered well at 24 h and locked tightly to display with a cobblestone-like appearance; the monolayer cells got confluence to 80%-90% after one-week submersion culture, and thereafter exposed to air-liquid interface. (2) Under scanning electron microscope, the cilia and also the small microvilli could be observed to protrude from the cell's surface at ALI day 7; the ciliated cells differentiated well and distributed in cluster; goblet cells and nonciliated columnar cells distributed between ciliated cells. (3) Immunocytochemistry of β-tubulin IV and zona occludens-1 showed a good confluence and differentiation of cilia in cultured epithelial cells at ALI day 14, and the percentage of ciliated epithelial cells was 50%-60 %. (4) The basal CBF of cultured epithelial cells was (8.42 ± 1.24), (8.71 ± 1.11), (9.17 ± 1.11), (8.89 ± 0.91), (8.99 ± 0.91) Hz at ALI day 7, 14 , 21, 28, 35, respectively, no significant difference was found among them(F = 1.451, P > 0.05). (5) At the concentration of 100 µmol/L ATP, an exogenous stimulating agent, significantly increased the CBF of cultured epithelial cells.

CONCLUSIONSAir-liquid interface cultured human nasal epithelial cells by enzymatic digestion method manifest with good confluence and differentiation status; the cells could maintain differentiated morphology and physiological function close to in vivo epithelium for a long term, therefore, it may serve as an ideal cell model for mucociliary transport study.

Cell Count ; Cell Culture Techniques ; Cells, Cultured ; Epithelial Cells ; cytology ; Humans ; Nasal Cavity ; cytology ; Nasal Polyps