Multicellul ar tumor spheroids of HeLa cells have been grown in a static culture system. Samples of spheroids were exposed for 2 h to graded concentration of sis-platinum and its analogue, carboplatin, and then response assayed by survival of clonogenic cells. The purpose of present experiment is to clarify the effectiveness of these platinum compounds and to evaluate intrinsic radiosensitivity of cells using spheroids of HeLa cells as an experimental in vitro model. Variations of the drug sensitivity of monolayers as well as spheroids were also evaluated in cell-survival curves. In cia-platinum concentration-survival cutie, there was a large shoulder extending as far as Cq=3.4 mM, after which there was exponential decrease in survival curve having a Co Value of 1.2 mu in spheroids. While the Co for the spheroids was essentially no significant change, but Cq value was larger than that of monolayers. This suggest that the effect of cis-platinum is greater in the monolayer with actively proliferaing cells than hypoxic one. In the carboplatin concentration-survival curves, the Co value of spheroids was 15.0 mM and the ratio with the Co from monolayer cell (32.5 mM) was 0.46, thus indicating that the spheroids had a greater sensitivity to carboplatin than monolayers. Therefore, the effect of carboplatin is mainly on the deeper layers of spheroids acting as hypoxic cell sensitizer. The enhanced effect was obtained for monolayer cells using combined X-ray and carboplatin treatment 2 hours before irradiation. The result shown in isobologram analysis for the level of surviving fraction at 0.01 indicated that the effect of two agents was truely supra-additive. From this experimental data, carboplatin has excited much recept interest as one of the most promising, since it is almost without nephrotoxicity and causes less gastrointestinal toxicity than cia-platinum. Interaction between carboplatin and radiation might play an important role for more effective local tumor control.
Carboplatin ; Cisplatin ; HeLa Cells* ; Humans ; Platinum Compounds ; Platinum* ; Radiation Tolerance ; Shoulder ; Spheroids, Cellular*

Hereditary diffuse leukoencephalopathy with neuroaxonal spheroids (HDLS) is a rare autosomal dominant leukoencephalopathy disease, and colony stimulating factor 1 receptor (CSF1R) is the only gene in which mutations are known to cause HDLS. HDLS should be suspected in individuals with progressive neurological decline, characteristic MR imaging findings, and positive family history. This article reviews recent advance in imaging findings, clinical manifestations, genetic counseling and management in HDLS.
Brain ; diagnostic imaging ; Humans ; Leukoencephalopathies ; diagnosis ; diagnostic imaging ; genetics ; physiopathology ; Radiography ; Spheroids, Cellular ; cytology

Three-dimensional (3D) culture of cells could closely mimic the in vivo situation with regard to cell function and microenvironment compared with plane monolayer cultured cells. In this paper, we established 3D culture of rat WB-F344 cells with rotary cell culture system (RCCS) to simulate microgravity environment, and examined cells proliferation, morphology, microstructure, E-cadherin protein quantity and mRNA expression of adhesion molecules by count the number of cells, optical microscope, transmission electron microscope and reverse transcriptase-polymerase chain reaction (RT-PCR). The results demonstrated that cells were polyhedron with lots of micovilli and mitochondria, which grow well and packed together densely to form irregular aggregates. Adjacent cells were connected with desmosome and tight junction. With the regard, the aggregates behaved 3D growth characteristics. Moreover, compared with control, mRNA level of Fibronectin and E-cadherin protein were increased, the changes maybe is the part mechanism in this microgravity simulated cells culture models which strengthened cells junction. This rotating 3D model might facilitate the study of interactions of cell-cell, cell-matrix and the mechanisms.
Animals ; Cadherins ; genetics ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cell Proliferation ; Fibronectins ; genetics ; Rats ; Spheroids, Cellular ; ultrastructure ; Weightlessness Simulation

When the size of a neurosphere cultured in vitro reaches a certain critical value, a necrotic core will appear inside the neurosphere because of the limitation of oxygen or other nutrients transport from medium to the cells in the neurasphere. Large necrotic core will greatly reduce the expansion of NSCs. The cellular automaton (CA) model is applied in this article to model the growth of NSCs in sphere state. The appearance and enlargement of the necrotic core in a neurosphere is calculated by coupling the CA model with the nutrient diffusion analysis in bioreactors. The calculation results indicate that the culture conditions, such as seeding density, the concentration of nutrients in medium and the mass transfer coefficient between a neurosphere and medium, have some effects on the appearance of the necrotic core. However, the necrotic core mainly depends on the inner diffusion. It will certainly appear if the size of the neurosphere is large enough even the outside mass transfer is in a good condition in bioreactors. Additionally, the appearance of the necrotic core resulting from the shortage of oxygen is earlier than that caused by the limitation of glucose. And the growth of the necrotic core is very fast after its appearance, and the whole neurosphere may become necrotic. The model developed with cellular automaton and mass transfer is a good qualitative representation of NSCs growth in bioreactors.
Bioreactors ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Computer Simulation ; Models, Biological ; Neurons ; cytology ; Spheroids, Cellular ; Stem Cells ; cytology ; Tissue Engineering ; methods

OBJECTIVETo imitate tumor microenvironment in vivo through construction of two-layer cell spheroid as a three-dimensional tumor model, and to validate its application in the study of Cx43 expression in colorectal cancer cell-fibroblast interactions and colorectal cancer progression.

METHODSThe two-layer cell spheroid was constructed from SW620 colorectal cancer cells and HELF fibroblasts. The expression of Cx43 in the spheroid was detected by immunocytochemistry. The expression of Cx43 in cultured SW620 cells with or without co-cultured fibroblasts was detected by immunocytochemistry and immunofluorescence. The expression of Cx43 in colorectal cancer tissue was detected by immunohistochemistry.

RESULTSThe spheroid showed well-defined cellular morphology and clear boundary between two cell lines.Significant expression of Cx43 was found along the boundary.SW620 cells had no expression of Cx43 when cultured alone, while the expression of Cx43 was induced upon co-culturing with fibroblasts.In the colorectal cancer tissue, expression of Cx43 was minimal in the centre of tumor in contrast to an upregulated expression at invasive front.

CONCLUSIONSThe two-layer cell spheroid is an observable and sensitive model for cell-cell interaction for studies of tumor microenvironment.It can simulate colorectal cancer cell-fibroblast interactions through up-regulation of Cx43 expression.

Cell Communication ; Cell Line, Tumor ; Coculture Techniques ; Colorectal Neoplasms ; metabolism ; pathology ; Connexin 43 ; metabolism ; Fibroblasts ; cytology ; Humans ; Spheroids, Cellular ; cytology ; metabolism ; Tumor Microenvironment ; Up-Regulation

OBJECTIVE: To prepare Arg-Gly-Asp (RGD) and cell penetrating peptide TAT co-modified paclitaxel loaded liposome (RGD/TAT-LP-PTX) for MCF-7 cell inhibition. METHODS: The co-modified liposome was prepared by film-ultrasonic method. The appearance, particle size and zeta potential were evaluated. The cellular uptake by MCF-7 cells in vitro was used to evaluate the targeting efficiency. The anti-proliferation efficiency of RGD/TAT-LP-PTX was evaluated by MTT assay. Tumor spheroids were used to evaluate anti-tumor ability of RGD/TAT-LP-PTX in vitro. RESULTS: The particle diameter of the co-modified liposome was (138.8 ± 12.4) nm with the Zeta potential of (25.85 ± 2.75) mV. The entrapment efficiency of PTX was 88.3%. The RGD/TAT-LP uptaken by MCF-7 cells at 4 h was 1.79 times higher than that at 2 h. The co-modified liposome uptaken by MCF-7 cells was 2.25 and 2.72 times higher than that of TAT-LP and RGD-LP, respectively. The anti-proliferation rate of RGD/TAT-LP-PTX increased with time. The inhibition rate of RGD/TAT-LP-PTX for MCF-7 cells at 48 h was 1.78 times higher than that at 24 h. The MTT assay demonstrated the cell viability of RGD/TAT-LP-PTX was 1.65, 1.82 and 2.55 times higher than that of TAT-LP-PTX, RGD-LP-PTX and LP-PTX, respectively. CONCLUSION Co-modified liposome may serve as a promising breast cancer delivery system for antitumor drugs.
Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; Cell Survival ; Humans ; Liposomes ; MCF-7 Cells ; drug effects ; Oligopeptides ; chemistry ; Paclitaxel ; pharmacology ; Particle Size ; Peptide Fragments ; chemistry ; Spheroids, Cellular ; drug effects

More than one-third of men may experience erectile dysfunction (ED) after nerve-sparing radical retropubic prostatectomy. A recent study has shown that vardenafil, a phosphodiesterase 5 inhibitor, could significantly improve the key indices of erectile function in men after unilateral or bilateral nerve-sparing radical retropubic prostatectomy. Few adverse events were observed in the study.
Double-Blind Method ; Erectile Dysfunction ; drug therapy ; etiology ; Humans ; Imidazoles ; therapeutic use ; Male ; Phosphodiesterase Inhibitors ; therapeutic use ; Piperazines ; therapeutic use ; Postoperative Complications ; drug therapy ; Prostatectomy ; Randomized Controlled Trials as Topic ; Spheroids, Cellular ; Sulfones ; therapeutic use ; Triazines ; therapeutic use ; Vardenafil Dihydrochloride

BACKGROUNDMonolayer cell culture models are the traditional culture models used for in vitro research of pancreatic carcinoma chemosensitivity. However, these models neglect the interactions between tumor cells and the impact of the tumor microenvironment. Such tumor cell monolayers poorly mimic the solid tumor microenvironment. The present study aimed to investigate the chemosensitivity characteristics of pancreatic cancer cells in a three-dimensional culture system by analyzing the differences in drug sensitivity between a scattered cell culture model and a multicellular spheroid culture model.

METHODSThree pancreatic cancer cell lines (SW1990, ASPC-1 and PCT-3) were cultured in three-dimensional collagen gels as well as in traditional two-dimensional monolayers. The chemosensitivities of the pancreatic carcinoma cells to 5-fluorouracil (5-FU), gemcitabine, and oxaliplatin in vitro were detected by both the Cell Counting Kit-8 test and the collagen gel droplet-embedded culture drug-sensitivity test.

RESULTSIn the two-dimensional culture model, differences in the chemosensitivities of the cloned pancreatic carcinoma cells and scattered cells existed for some concentrations of 5-FU, gemcitabine and oxaliplatin. In the three-dimensional culture model, there were significant differences in the chemosensitivities of the pancreatic cancer cells between the scattered cells and multicellular spheroids (P < 0.05).

CONCLUSIONPancreatic carcinoma cells exhibit multicellular resistance in three-dimensional cultures.

Antineoplastic Agents ; pharmacology ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Cell Survival ; drug effects ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Fluorouracil ; pharmacology ; Humans ; Organoplatinum Compounds ; pharmacology ; Pancreatic Neoplasms ; drug therapy ; Spheroids, Cellular ; drug effects

OBJECTIVE: To investigate the relationship between matrix matrix metalloproteinases (MMP)-9 expression in HCE1/multicellular spheroids (MCS) and invasion of cervical carcinoma, and to explore the effect of MMPs inhibitor GM6001 on the model that HCE1/MCS invade live human umbilical vein endothelium cell (HUVEC). METHODS: We established the invasion model by coculturing HCE1/MCS and HUVEC, and detected the MMP-9 expression in HCE1 monolayer cells (HCE1/MC), HCE1/MCS and HUVEC using immunohistochemistry and treated the HCE1/MCS invasion model with GM6001. RESULTS: A cervical squamous carcinoma cell HCE1 infiltrating model was established. Compared with that in HCE1/MC, MMP-9 expression in the HCE1/MCS was significantly higher (P<0.05). Compared with the control group, the invasion of HCE1/MCS was inhibited by 26.09% and 92.95% (P<0.05) in the GM6001 2.5 μmol/L and 12.5 μmol/L group. MMP-9 expression in HCE1/MCS in the GM6001 12.5 μmol/L group was obviously lower than that in the control group (P<0.05). With the increase of the concentration of GM6001, the inhibition increased. CONCLUSION The enhancement of HCE1/MCS invasion may be related to the up-regulation of MMP-9 expression in HCE1/MCS. GM6001 can down regulate the MMP-9 expression in HCE1/MCS and partially block the HCE1/MCS invasion to HUVEC.
Carcinoma, Squamous Cell ; pathology ; Cell Line ; Cell Line, Tumor ; Dipeptides ; pharmacology ; Endothelial Cells ; pathology ; Female ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Spheroids, Cellular ; metabolism ; pathology ; Umbilical Veins ; cytology ; Uterine Cervical Neoplasms ; pathology

To investigate the effect of manganese superoxide dismutase (MnSOD) silence on the in vitro tumorigenicity in human small cell lung cancer NCI-H446 cells and the underlying mechanisms.
 Methods: Sphere formation cells from NCI-H446 cells were obtained by suspension culture, while the expression of MnSOD and urokinase type plasminogen activator (uPAR) was analyzed by Western blot. Silence of MnSOD was performed by adenovirus infection in the second passage formation cells, and the effect of MnSOD silence on tumorigenicity in NCI-H446 cells was evaluated by sphere formation assay and soft-agar colony formation assay, while the expression of uPAR was analyzed by Western blot.
 Results: Compared with NCI-H446 cells, the sphere formation rate, colony formation rate, and the expression of MnSOD and uPAR were significantly increased in the second passage sphere formation cells in NCI-H446 cells (P<0.05). Silence of MnSOD inhibited the sphere formation rate, colony formation rate, and the expression level of uPAR in the second passage sphere formation cells in NCI-H446 cells.
 Conclusion: MnSOD may promote tumorigenicity in NCI-H446 cells by up-regulation of uPAR expression in vitro.
Adenoviridae ; Carcinogenesis ; Cell Line, Tumor ; Humans ; In Vitro Techniques ; Lung Neoplasms ; etiology ; metabolism ; RNA Interference ; Receptors, Urokinase Plasminogen Activator ; genetics ; metabolism ; Small Cell Lung Carcinoma ; etiology ; metabolism ; Spheroids, Cellular ; pathology ; Superoxide Dismutase ; genetics ; metabolism ; Tumor Stem Cell Assay ; Up-Regulation