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

OBJECTIVETo investigate the biological characteristic of ectomesenchymal stem cells (EMSC) and the differentiation of p75(+)EMSC and influencing factors.

METHODSThe immunohistochemical staining method was used to observe the migration of EMSC from 12.5 d SD rat embryonic facial process. Then EMSC was labeled by p75 neurotrophin receptor, and the cell cycle and stem cell surface antigens of the p75(+) EMSC was examined.

RESULTSImmunohistochemical staining showed that stem cells from the cranial neural crest migrated to embryonic rat facial process at 12.5 days. The sorting rate of the p75(+)EMSC was 6.1%. The proportion of the p75(+)EMSCs' S/G2/M phase was stable during subculture. The special substances CD29, CD146 and Stro-1 were marked for p75(+)EMSC, and the expressions of the markers were all higher (> 90%) in p75(+) EMSC.

CONCLUSIONSThe embryonic tooth did not start to grow on the conception 12.5 days of SD rats. The p75(+) EMSC after sorting had stable proliferation ability and had stem cell characteristics during subculture and didn't start differentiation.

Animals ; Biomarkers ; metabolism ; Cell Cycle ; Cell Differentiation ; Cell Division ; Cell Movement ; In Vitro Techniques ; Mesenchymal Stromal Cells ; chemistry ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptor, Nerve Growth Factor

OBJECTIVES: The etiology for Kawasaki disease (KD) remains unknown, but several studies have suggested the involvement of immune dysregulation and genetic factors. The purpose of this study is to compare gene expressions before and after an infusion of intravenous immunoglobulin (IVIG) in KD patients. METHODS: Blood was obtained from both acute and sub-acute phases of 4 patients with KD and febrile control children. Blood was collected in PAXgene blood RNA tubes and RNA was extracted using a PAXgene blood RNA isolation kit. Labeled RNAs were analyzed using Roche NimbleGen human whole genome 12-plex array. RESULTS: KD patients prior to IVIG injection showed more than a two-fold increase in the expression of 88 genes and more than a two-fold decrease in the expression of 98 genes compared to the control group. They also showed more than two-fold increase in the expression of 226 genes and more than a two-fold decrease in 117 genes in KD patients after IVIG treatment compared to the patients before IVIG injection. Through microarray evaluation, the expressions of genes involved in proliferation, translation, inflammatory response, immune response, cell adhesion, cell migration, cell differentiation, apoptosis, cell growth, transport, cell cycle, transcription, signal transduction and metastasis were observed. CONCLUSION: Changes in gene expressions in pediatric patients with KD before and after IVIG were observed via microarray evaluation.
Apoptosis ; Cell Adhesion ; Cell Cycle ; Cell Differentiation ; Cell Movement ; Child ; Gene Expression ; Genome ; Humans ; Immunoglobulins ; Immunoglobulins, Intravenous ; Microarray Analysis ; Mucocutaneous Lymph Node Syndrome ; Neoplasm Metastasis ; RNA ; Signal Transduction

OBJECTIVETo investigate the effects of RCCS on cell seeding onto 3-D scaffold and cell-scaffold composite culture in vitro.

METHODSRabbit skin fibroblasts of passage 2 were seeded at 2 x 10(6) cell per cm(3) onto/into polyglycolic acid (PGA) foams by static seeding (dropping a cell suspension onto foams) or dynamic seeding (rotating PGA foams and a cell suspension in RCCS). Attachment of cells in foams was observed by cell-counting after trypsin digestion. The effects of culture condition were next studied by culturing cell-PGA complexes in RCCS versus static culture condition. Distribution and proliferation of cells in foams were investigated with MTT, stereomicroscope and scan electron microscope.

RESULTSNumbers of cells adhering to polymers increased gradually during an initial period of 24 hours. Eight, 12 and 24 hours after seeding, the rates of adhering cells were significantly higher in the dynamic seeding group than in the static seeding group (46.70% + 2.16% vs. 31.50% +/- 3.54%; 56.36% +/- 3.18% vs. 34.28% +/- 3.16%; 66.32% +/- 4.60% vs. 37.38% +/- 4.66%; P < 0.01). The dynamic culture method as compared to the static method resulted in new tissues with a higher cellularity and more uniform cell distribution during a 3 period of weeks.

CONCLUSIONSRCCS has advantages of promoting cell attachment, uniform migration and proliferation in polymer scaffolds and can be used for construction of 3-D cell-polymer tissues in vitro.

Animals ; Cell Adhesion ; Cell Culture Techniques ; methods ; Cell Division ; Cell Movement ; Fibroblasts ; cytology ; Polyglycolic Acid ; pharmacology ; Rabbits ; Time Factors

P21-activated kinases (PAK) participate in a variety of important cellular activities, such as cytoskeleton remodeling, cell migration, cell cycle regulation, and apoptosis or survival. PAK also has an important impact on brain development, neuronal differentiation, and regulation of synaptic plasticity in the nervous system. PAK abnormalities result in diseases including cancer, Parkinson's disease (PD), Alzheimer's disease (AD) and neural retardation. Therefore, it is of vital physiological significance to investigate the neuronal function of PAK. In this paper we review the advancement of research on the neuronal biological function and the underlying mechanisms of PAK.
Alzheimer Disease ; physiopathology ; Apoptosis ; Cell Cycle ; Cell Movement ; Cytoskeleton ; physiology ; Humans ; Nervous System ; enzymology ; Neuronal Plasticity ; Neurons ; physiology ; Parkinson Disease ; physiopathology ; p21-Activated Kinases ; physiology

Endothelial injury, smooth muscle cells (SMCs) proliferation and migration are the same common pathophysiological processes of many cardiovascular diseases, such as atherosclerosis, hypertension, diabetes and restenosis. It is important to determine the functional interactions between endothelial cells (ECs) and SMCs under pathologic conditions. This work was to study the effects of ECs growth states on the proliferation and migration of vascular SMCs in cell coculture system. (3)H-TdR incorporation and flow cytometry were used to determine the effects of ECs growth states on the proliferation of SMCs. The number of migrating SMCs was counted. RT-PCR was used to analyze the expression of alpha-SM-actin mRNA. The results showed that (3)H-TdR incorporation decreased significantly from 14,900+/-1035 cpm/well in the control group to 8,575+/-749 cpm/well in the confluent ECs group (n=6, P<0.01), and increased to 27,268+/-2310 cpm/well in the proliferative ECs group ( n=6, P<0.01). The transition of SMCs from G(0)/G(1) phase to G(2)/M and S phases was blocked in the confluent ECs group but promoted in the proliferative ECs group. Compared with the control group, the number of migrating cells was about 4 times higher in the proliferative ECs group (n=6, P<0.01), while it in the confluent ECs group was only the half of the number of the control(n=6, P<0.05). The expression of alpha-SM-actin mRNA was increased significantly in the confluent ECs group(2.3+/-0.11 vs 1.4+/-0.12, P<0.05), but decreased significantly in the proliferative ECs group(0.92+/-0.08 vs 1.4+/-0.12, P<0.05). The results suggest that the biologic features of SMCs are influenced by ECs growth states. The proliferative ECs promote SMCs proliferation, migration and downregulate alpha-SM-actin mRNA expression significantly.
Actins ; metabolism ; Aorta ; cytology ; Cell Differentiation ; Cell Division ; Cell Movement ; Cells, Cultured ; Coculture Techniques ; Endothelium, Vascular ; cytology ; Epithelial Cells ; cytology ; Muscle, Smooth, Vascular ; cytology

To evaluate the feasibility of using human embryonic fibroblast(HEF) as feeder layer in the culture of human embryonic stem(ES) cells in vitro, we investigated the morphology, the sensitivity to 0.25% trypsin, the growth curve and cell cycle of HEF with DMEM(low glucose) +10% FBS used as culture medium, and then we compared HEF with mouse embryonic fibroblast (MEF). The results showed that both HEF and MEF are adherent cells in vitro, and HEF has longer life span and better growth ability than MEF. In room temperature, HEF is more sensitive to 0.25% trypsin. Our research suggested that HEF can be used as feeder layer in culture of ES cells. HEF has longer service life than MEF and is worthy to be studied further.
Animals ; Cell Cycle ; Cell Differentiation ; Cell Division ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Fibroblasts ; cytology ; physiology ; Humans ; Mice

OBJECTIVETo study the adhesion, migration and three-dimentional growth of Schwann cells on PLA (polylactic acid) nonspinning fibre cloth and polyglycolic/polylactic acid (PLGA) fibres.

METHODSSchwann cells/ECM gel solution and PLA nonspinning fibre cloth and PLGA fibres pretreated by collagen, polylysine and ECM were co-cultured. Then the migration and three-dimensional growth of Schwann cells on the fibres were observed under phase contrast microscope and laser scanning confocal microscope.

RESULTSSchwann cell/ECM solution was compounded with PLA nonspinning fibre cloth. With formation of gel, most Schwann cells resided in the fibre net holes, and adhered to the fibres to form a multiplayer-arranged Schwann cell column like Büngner band. Schwann cells could adhere to PLGA fibres and grew and migrated along the fibres. ECM gel could significantly increase the adhering and migrating cell number.

CONCLUSIONSECM gel can facilitate the adhesion, growth and migration of Schwann cells on the seteroframe. It is a good integrating material for tissue engineering bioartificial nerve.

Cell Adhesion ; Cell Division ; Cell Movement ; Humans ; Lactic Acid ; Polyglycolic Acid ; Polymers ; Schwann Cells ; physiology ; Tissue Engineering

BACKGROUND: The structure and function of bone tissue is maintained through a constant remodeling process, which is maintained by the balance between osteoblasts and osteoclasts. The failure of bone remodeling can lead to pathological conditions of bone structure and function. Remifentanil is currently used as a narcotic analgesic agent in general anesthesia and sedation. However, the effect of remifentanil on osteoclasts has not been studied. Therefore, we investigated the effect of remifentanil on pre-osteoclast (pre-OCs) differentiation and the mechanism of osteoclast differentiation in the absence of specific stimulus. METHODS: Pre-OCs were obtained by culturing bone marrow-derived macrophages (BMMs) in osteoclastogenic medium for 2 days and then treated with various concentration of remifentanil. The mRNA expression of NFATc1 and c-fos was examined by using real-time PCR. We also examined the effect of remifentanil on the osteoclast-specific genes TRAP, cathepsin K, calcitonin receptor, and DC-STAMP. Finally, we examined the influence of remifentanil on the migration of pre-OCs by using the Boyden chamber assay. RESULTS: Remifentanil increased pre-OC differentiation and osteoclast size, but did not affect the mRNA expression of NFATc1 and c-fos or significantly affect the expression of TRAP, cathepsin K, calcitonin receptor, and DC-STAMP. However, remifentanil increased the migration of pre-OCs. CONCLUSIONS: This study suggested that remifentanil promotes the differentiation of pre-OCs and induces maturation, such as increasing osteoclast size. In addition, the increase in osteoclast size was mediated by the enhancement of pre-OC migration and cell fusion.
Anesthesia, General ; Bone and Bones ; Bone Remodeling ; Cathepsin K ; Cell Differentiation ; Cell Fusion ; Cell Movement ; In Vitro Techniques ; Macrophages ; Osteoblasts ; Osteoclasts ; Real-Time Polymerase Chain Reaction ; Receptors, Calcitonin ; RNA, Messenger

Chitosan is a natural biopolymer and is made up of D-glucosamine subunits linked by beta-(1,4) glycosidic bond. In recent years, the application of chitosan has attracted more and more attention because of its good biological function in cell biology. The properties of chitosan-based biomaterial are attributed to the physical properties and chemical composition of chitosan. The author of this paper summarized recent related studies and progresses of the influence of physical properties of chitosan on cell activity and cell mechanics property at home and abroad. The findings show that most studies mainly focused on the influence of chitosan and cell activity, while few were on cell mechanics property. The related studies of the influence of chitosan on cell will contribute to the explanation for the mechanism of the interaction between chitosan and cell, and provide the theoretical support for the further study.
Animals ; Cell Adhesion ; drug effects ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Chemical Phenomena ; Chitosan ; chemistry ; pharmacology ; Humans ; Tissue Engineering ; Tissue Scaffolds ; chemistry