Eccrine porocarcinoma (EP) is a rare malignant tumor arising from the intraepidermal eccrine duct. The tumor cells frequently contain glycogen, but prominent clear cell changes in EP are rarely reported. A 78-year-old woman presented with a slightly pruritic, erythematous, verrucous plaque on her left thigh. Histopathological examination revealed intraepidermal tumor cell nests composed of small basaloid cells and duct-like structures lined by periodic acid-Schiff (PAS)-positive cuticles. Besides the typical findings of EP, clear cell changes were predominantly observed in the tumor cell aggregations. Herein we report a case of the clear cell variant of EP rarely reported in previous literature.
Aged ; Cell Aggregation ; Eccrine Porocarcinoma ; Female ; Glycogen ; Humans ; Thigh

By using the size distribution of cell aggregates, viable cell density, cell viability, specific consumption rate of glucose (q(glc)), specific production rate of lactate (q(lac)) and lactate transform rate (Y(lac/glc)) as the evaluation indexes, the effects of hydrodynamic on aggregates formation, growth and metabolism of HEK293 cells in suspension culture were examined in 250mL spinner-flasks by setting the agitation rates at 25, 50, 75 and 100r/min, respectively. It was found that agitation plays an important role in HEK293 cell aggregates formation and cell aggregates size distribution. After 7d cultivation in spinner-flasks operated at 50r/min and 75r/min, the average diameter of HEK293 cell aggregates was 201 microm and 175 microm, respectively, with the fraction of aggregates larger than 225 microm less than 10%. The cell viability was kept above 90% with the metabolic indexes, including q(glc), q(lac) and Y(lac/glc) kept constant. These results demonstrated that hydrodynamic derived from the proper agitation play a decisive role in controlling the formation and size distribution of HEK293 cell aggregates, and provided sufficient mass transfer to support the normal growth and metabolism of HEK293 cells in suspended aggregates.
Bioreactors ; Cell Aggregation ; Cell Culture Techniques ; instrumentation ; Cell Line ; Cell Proliferation ; Humans ; Kinetics

Human epithelial cell line immortalized by Ad12-SV40 hybrid virus was transfected with plasmid containing HPV-16 via calcium-phosphate method. Subsequently, 8 clonal cell lines were obtained after G418 selection. Among these clonal cells, clonal cell-4(C-4) and clonal cell-6(G-6) showed increases of tumorigenic cellular properties such as saturation density, soft agar colony formation and cell aggregation. Morphological alteration such as appearane of foci was observed on these two clones after passage 6 and 7(50 to 55 days after transfection). When clonal cells and control cells were treated with MNNG(0.01ug/ml), both C-4 and C-6 showed increases of tumorigenic cellular properties and the level of increase was much more elevated as compared to clonal cells prior to MNNG treatment. Appearance of foci formation was observed in C-4 and C-6 after passage-6. After passage-8, control cells and all clonal cells showed morphological alterations. It seems that treatment of cells containing HPV-16 DNA with MNNG increases tumorigenic properties of the cells and expedite morphological transformations. (continue)
Agar ; Cell Aggregation ; Cell Line ; Clone Cells ; DNA ; Epithelial Cells* ; Human papillomavirus 16 ; Humans* ; Methylnitronitrosoguanidine ; Plasmids

The investigation of the mechanism of biological pattern has been an important topic of life sciences, especially, of developmental biology, for a long time. It is an interdisciplinary problem and many researching data have been obtained and some theories have been structured from many points of view in science. However, up to now, the actual mechanism is still a fascinating puzzle and needs more studies. In this paper, we try to construct a cellular automata model of biological pattern. This model defines the individual model cells and their behaviors, cell-cell interactions, and cell-environment interactions. As an application, we present a new discrete model to simulate the aggregation phase of the development of Dictyostelium discoideum with the concept of "inducing switch".
Animals ; Cell Aggregation ; physiology ; Cell Movement ; physiology ; Dictyostelium ; cytology ; physiology ; Models, Biological

OBJECTIVE: To explore whether the differentiation of vascular stem cells (VSC) into smooth muscle cells (SMC) in aortic dissection (AD) is dysregulated, and to verify the role of Notch3 pathway in this process. METHODS: Aortic tissues were obtained from AD patients undergoing aortic vascular replacement and heart transplant donors at Department of Cardiovascular Surgery, Guangdong Provincial People's Hospital Affiliated to Southern Medical University. VSC were isolated by enzymatic digestion and c-kit immunomagnetic beads. The cells were divided into normal donor-derived VSC group (Ctrl-VSC group) and AD-derived VSC group (AD-VSC group). The presence of VSC in the aortic adventitia was detected by immunohistochemical staining, and VSC was identified by stem cell function identification kit. The differentiation model of VSC into SMC established in vitro was induced by transforming growth factor-β1 (10 μg/L) for 7 days. They were divided into normal donor VSC-SMC group (Ctrl-VSC-SMC group), AD VSC-SMC group (AD-VSC-SMC group) and AD VSC-SMC+Notch3 inhibitor DAPT group (AD-VSC-SMC+DAPT group,DAPT 20 μmol/L was added during differentiation induction). The expression of contractile marker Calponin 1 (CNN1) in SMC derived from aortic media and VSC were detected by immunofluorescence staining. The protein expressions of contractile markers α-smooth muscle actin (α-SMA), CNN1 as well as Notch3 intracellular domain (NICD3) in SMC derived from aortic media and VSC were detected by Western blotting. RESULTS: Immunohistochemical staining showed there was a population of c-kit-positive VSC in the adventitia of aortic vessels, and VSC from both normal donors and AD patients had the ability to differentiate into adipocytes and chondrocytes. Compared with normal donor vascular tissue, the expressions of SMC markers α-SMA and CNN1 of tunica media contraction in AD were down-regulated (α-SMA/β-actin: 0.40±0.12 vs. 1.00±0.11, CNN1/β-actin: 0.78±0.07 vs. 1.00±0.14, both P < 0.05), while the protein expression of NICD3 was up-regulated (NICD3/GAPDH: 2.22±0.57 vs. 1.00±0.15, P < 0.05). Compared with Ctrl-VSC-SMC group, the expressions of contractile SMC markers α-SMA and CNN1 were down-regulated in AD-VSC-SMC group (α-SMA/β-actin: 0.35±0.13 vs. 1.00±0.20, CNN1/β-actin: 0.78±0.06 vs. 1.00±0.07, both P < 0.05), the protein expression of NICD3 was up-regulated (NICD3/GAPDH: 22.32±1.22 vs. 1.00±0.06, P < 0.01). Compared with AD-VSC-SMC group, the expressions of contractile SMC markers α-SMA, CNN1 were up-regulated in AD-VSC-SMC+DAPT group (α-SMA/β-actin: 1.70±0.07 vs. 1.00±0.15, CNN1/β-actin: 1.62±0.03 vs. 1.00±0.02, both P < 0.05). CONCLUSIONS Dysregulation of VSC differentiation into SMC occurs in AD, while inhibition of Notch3 pathway activation can restore the expression of contractile proteins in VSC-derived SMC in AD.
Humans ; Actins ; Platelet Aggregation Inhibitors ; Signal Transduction ; Aortic Dissection ; Cell Differentiation ; Myocytes, Smooth Muscle ; Stem Cells

OBJECTIVE: Recently it has been proposed that stem cells may be associated with the pathogenesis of endometriosis. The purposes of this study are to investigate whether the eutopic endometrial cells of women with or without endometriosis show the characteristics of stem cells in vitro and have a difference of the expressions of the undifferentiated stem cell markers as OCT-4 and CXCR4. METHODS: A total of 6 women with advanced endometriosis and a total of 10 women without endometriosis, adenomyosis or leiomyoma were included in this study. The eutopic endometrial cells, which were obtained from the menstrual blood at menstrual cycle day 2 to 4, were cultured in vitro for approximately 2 weeks, subsequently the putative very small stem cells were separated by Percoll density gradient method and were cultured. The expressions of OCT-4 and CXCR4 were analyzed by real time RT-PCR. RESULTS: The eutopic endometrial cells of the group of endometriosis compared with the control group showed the different morphological characteristics in vitro; more commonly heterogeneous supportive cells, very small round cells less than 3 micrometer and 5~15 micrometer sized hyperchromatic round cells. After the separation of very small round cells by Percoll density gradient method, these cells showed the several characteristics of stem cells; self-renewal, asymmetric cell division, colony formation and embryoid body-like formation. Also These cells showed the similar characteristics of very small embryonic-like stem cells; the mobile cells smaller than erythrocyte, the cell migration or adhesion to supportive cells, the sphere formation by cell aggregation and the formation of new differentiated cell by cell fusion. The expressions of OCT-4 and CXCR4 in the group of endometriosis are respectively 5.66 times and 17.69 times as high as the control group (P<0.05). CONCLUSION: The very small round cells less than 3 micrometer and 5~15 micrometer sized hyperchromatic round cells, which showed the several characteristics of stem cells in vitro, were more common in eutopic endometrial cells of patients with endometriosis and the expressions of OCT-4 and CXCR4 were significantly higher. This study suggests that stem cells might play a key role in the pathogenesis of endometriosis and OCT-4 and CXCR4 might be used as a tool for diagnosis or follow-up.
Adenomyosis ; Asymmetric Cell Division ; Cell Aggregation ; Cell Fusion ; Cell Movement ; Endometriosis ; Endometrium ; Erythrocytes ; Female ; Humans ; Leiomyoma ; Menstrual Cycle ; Povidone ; Silicon Dioxide ; Stem Cells

The effect of calcium on the aggregation and growth of 293 cells grown in either serum-containing or serum-free medium was investigated in T-flask and spinner bottle, respectively. It was found that the concentration of calcium ion, in the range of 0.1 mmol/L to 1.0 mmol/L, affected adhesion and aggregation of 293 cells severely, but had no distinct effect on growth. The result indicated that the attachment of 293 cells was easier with higher calcium concentration in serum-containing medium. And 293 cells formed aggregates readily in suspension culture. This effect was more profound in cultures with higher calcium concentration. The average diameter( D, mum) of 293 cell aggregates exhibited direct proportion to calcium concentration (c, mmol/L) in serum-free medium. It can be depicted by a simple equation in the range of 0.1 mmol/L to 0.5 mmol/L, i.e. D = 58.65c + 16.96. The aggregation size of 293 cells is regulable in suspension culture, therefore, proper control allows for an easier cell retention, and thus a high cell concentration potentially can be achieved. However, similar growth of 293 cells was observed in cultures with different calcium concentration.
Calcium ; pharmacology ; Cell Aggregation ; drug effects ; Cell Culture Techniques ; Cell Division ; drug effects ; Cell Line ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Humans

Lectins are glycoproteins that bind specifically to carbohydrates. Considerable interests in the lectins were encouraged by several reports that certain members of the family bind to the extracellular matrix proteins (ECM), such as fibronectin and laminin. However, the relations between lectin and ECM protein remain unclear. To elucidate the relations of lectin-matrix-cell, we treated three cancer cell lines, HeLa, L929, and EATC with ConA and PHA-P at low dose (4 microgram/ml) and high dose (20 microgram/ml) for 1, 3, 5 days. 1. Whether or not lectins significantly regulate the cell proliferation was evaluated by MTT assays. 2. Whether the amount of fibronectin and laminin which of cancer cells can be influenced by lectins was confirmed by immunocytochemical staining. 3. Whether, in turn, the lectins which can change the morphology were observed under inverted and electron microscopes. ConA and PHA-P inhibited cell proliferation rate of all cell lines in a dose- and time- dependent manner. The amount of fibronectin and laminin considerably reduced in the three cell lines after the lectins treatment in a dose- and time-dependent manner. The cancer cell lines showed various morphological changes such as cell aggregation, irregular-shaped cellular processes, rounded cells, cytoplasmic vacuolation, swollen RERs, dilation of mitochondria, margination of chromatin and cell death. In conclusion, our results showed ConA and PHA-P caused damages of the three cancer cell lines, but the effect of PHA-P was much stronger than ConA. Taken together, the present data strongly indicate that ConA and PHA-P influence the cell proliferation rate, reduce the amount of fibronectin and laminin and induce cell injuries of HeLa, L929, and EATC cell lines. Our results also suggest that the cancer cell proliferation and the morphological changes might be modulated by the specific interaction between lectins and ECM proteins associated with the cell surface.
Carbohydrates ; Cell Aggregation ; Cell Death ; Cell Line* ; Cell Proliferation ; Chromatin ; Cytoplasm ; Extracellular Matrix Proteins ; Extracellular Matrix* ; Fibronectins ; Glycoproteins ; Humans ; Laminin ; Lectins ; Mitochondria

BACKGROUND: Although apoptosis occurs in nucleated cells, studies show that this event also occurs in some anucleated cells such as platelets. During storage of platelets, the viability of platelets decreased, storage lesions were observed, and cells underwent apoptosis. We investigated the effects of caspase-3 inhibitor on the survival and function of platelets after different periods of storage. METHODS: Platelet concentrates were obtained from the Iranian Blood Transfusion Organization in plastic blood bags. Caspase-3 inhibitor (Z-DEVD-FMK) was added to the bags. These bags along with control bags to which no inhibitor was added were stored in a shaking incubator at 22degrees C for 7 days. The effects of Z-DEVD-FMK on the functionality of platelets were analyzed by assessing their ability to bind to von Willebrand factor (vWF) and to aggregate in the presence of arachidonic acid and ristocetin. Cell survival was surveyed by MTT assay. RESULTS: At day 4 of storage, ristocetin-induced platelet aggregation was significantly higher in the inhibitor-treated (test) than in control samples; the difference was not significant at day 7. There was no significant difference in arachidonic acid-induced platelet aggregation between test and control samples. However, at day 7 of storage, the binding of platelets to vWF was significantly higher in test than in control samples. The MTT assay revealed significantly higher viability in test than in control samples at both days of study. CONCLUSION: Treatment of platelets with caspase-3 inhibitor could increase their functionality and survival.
Apoptosis ; Arachidonic Acid ; Blood Platelets* ; Blood Transfusion ; Caspase 3* ; Cell Survival ; Incubators ; Plastics ; Platelet Aggregation ; Ristocetin ; von Willebrand Factor

This study was aimed to investigate the effect of prefreezing parameters such as freezing rate, annealing, rate, annealing temperature, holding time in annealing before lyophilization on lyophilized platelets by orthogonal tests. The recovery rate, the morphological and ultrastructural changes, the activation and aggregation of lyophilized platelet after rehydratation were detected by cytometer, scan electron microscopy, flow cytometry and aggregation reaction on thrombin, respectively, and complex evaluation was carried out. The results showed that the recovery rate of lyophilized platelets was 91% - 53.5% at different conditions of lyophilization, the size and shape of ice crystals in lyophilized platelets of different test groups were not similar, the expression and distribution of platelet activation markers (PAC-1 and CD62p) after rehydration were relatively similar to fresh platelets, expression rate of PAC-1 was lower (0.03% - 0.22%), meanwhile there were differences of CD62p expression levels between different groups. The optimal theoretic composition of prefreezing conditions obtained on basis of platelet recovery rate was A(2)B(1)C(1)D(3), i.e, first, suspensions of platelets on the program control cooling apparatus were lyophilized with a rate of 20 degrees C/min and maintained at -40 degrees C for 2 hours, then the annealing was conducted at -30 degrees C for 0.5 hours with a heating rate of 1.5 degrees C/min; finally, the freeze-drying program was carried out to the end. It is concluded that the freezing rate, annealing rate, annealing temperature and holding time of annealing all impact on the recovery of lyophilized platelets. The preservation qualities of lyophilized platelets were affected by the various combinations of prefreezing parameters.
Blood Platelets ; Blood Preservation ; methods ; Cell Survival ; Cryopreservation ; methods ; Freeze Drying ; methods ; Humans ; Platelet Activation ; Platelet Aggregation ; Platelet Count