OBJECTIVETo investigate the biological characteristics of cell lines of healthy and diseased human dental alveoli.

METHODSPrimary cell lines from either healthy or diseased human dental alveoli were obtained. Two cell lines, H-258 and H-171 derived from healthy and diseased human tissues respectively, were selected for morphological study and research on their growth and aging, using cell counting, and histochemical and immunohistochemical staining.

RESULTSPrimary cell lines were successfully established from innormal dental alveoli. After freezing and thawing for three times, cell growth was continued and no morphological alterations were observed. The doubling time was 53.4 hours and mean division index (MDI) was 4 per thousand. Cells were kept normal after twenty generations with no obvious reduction of doubling time and MDI. Of twenty-six primary cell lines derived from healthy human dental alveoli, only three cell lines achieved generation. After freezing and thawing for twice, cultured cells were still alive at a decreased growth speed, with doubling time of 85.9 hours and MDI of 3 per thousand. Both cell lines, H-171 and H-258, shared the characteristics of osteoblast.

CONCLUSIONSPrimary cell lines of diseased human dental alveoli show greater growth potential. All cell lines of dental alveoli share characteristics of osteoblast. The technique we developed may be put into practice for the treatment of abnormal dental alveoli.

Cell Division ; physiology ; Cell Line ; Humans ; Tooth Socket ; cytology

This study was aimed to investigate the biological characteristics of osteoblasts and their hematopoietic supportive function by using human fetal osteoblastic cell line 1.19 (hFOBs) as a model. The pluripotency markers (Oct-4, Rex-1, hTERT) of hFOBs were analyzed by RT-PCR, the multilineage differentiation experiments were conducted in vitro. Flow cytometry (FCM) was used to identify the surface markers of hFOBs, and RT-PCR was used to analyze their hematopoietic cytokine expression in comparison with bone marrow mesenchymal stem cell (BM-MSC). The results showed that hFOBs expressed several ESC pluripotency markers including Oct-4 and Rex-1, except hTERT. Moreover, hFOBs could also undergo multilineage differentiation into the mesodermal lineages of adipocytic cell types in addition to its predetermined pathway, the mature osteoblast. Both hFOBs and BM-MSC expressed CD44, CD73 (SH3), CD105 (SH2) and CD90 (Thy1), and lack expression of CD34, CD45, or HLA-DR surface molecules. In addition, both hFOBs and BM-MSC expressed SCF, IL-6, and SDF-1alpha mRNA, but only hFOBs could express GM-CSF and G-CSF. It is concluded that human fetal osteoblastic cell line 1.19 may provide a good model to study the osteoblastic regulation role in hematopoiesis in vitro.
Cell Differentiation ; physiology ; Cell Line ; Fetus ; Hematopoiesis ; physiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology ; Models, Biological ; Osteoblasts ; cytology ; physiology

To investigate the effects of human mesenchymal stem cells (MSC) and human fibroblastoid cell line (HFCL) as feeder layer on expansion of umbilical cord blood CD34(+) cells in vitro, (60)Co gamma-ray irradiated MSC and HFCL were used as feeder layer to expand cord blood CD34(+) cells in culture. The efficiencies of MSC and HFCL on expansion of CD34(+) cells in culture with or without cytokines were compared. The results showed that no matter whether cytokines (rhFL, rhSCF, rhTPO) were added, the proliferation of nucleated cells after expansion for 12 days in HFCL group was statistically higher than that in MSC group, i.e. with cytokines (9797 +/- 361)% vs (7061 +/- 418)%; without cytokines (5305 +/- 354)% vs (1992 +/- 247)%, when the cell numbers at day 0 was accounted as 100%), P < 0.01. The proliferation of propagated CD34(+) cells between MSC group and HFCL without addition of cytokines was not statistically different (820 +/- 191)% vs (825 +/- 305)%, P > 0.05. However, in the presence of cytokines, the propagating rate of MSC group was lower than that of HFCL group (939 +/- 212)% vs (1617 +/- 222)%, P < 0.01. MSC was better than HFCL in maintaining the LTC-IC of UCB CD34(+) cells, i.e. the number of CFU-GM colonies in the fifth week was (129.95 +/- 8.73) /10(5) seeded cells vs (89.81 +/- 10.29) colonies/10(5) cells, P < 0.05; with addition of cytokines, the effect was more obvious, i.e. the number of CFU-GM colonies in the fifth week (192.93 +/- 4.95)/10(5) seeded cells vs (90.47 +/- 14.28) colonies/10(5) seeded cells, P < 0.01. MSC mixed with a certain proportion of HFCL facilitated maintaining the LTC-IC of UCB CD34(+) cells. When the proportion was 4:1, the number of CFU-GM colonies was the highest (186.89 +/- 11.11)/10(5) seeded cells, which was higher than that of both 3:2 group [(138.92 +/- 14.84) colonies/10(5) seeded cells] and MSC only group, i.e. (64.63 +/- 6.11) colonies/10(5) seeded cells, both P < 0.01. It is concluded that HFCL is better than MSC in maintaining the expansion of CD34(+) cells and cytokines can enhance this effect, while MSC are stronger than HFCL in maintaining the LTC-IC of UCB CD34(+) cells in vitro. MSC with addition of a certain proportion of HFCL can significantly enhance the efficiency of CD34(+) cell expansion.
Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; physiology ; Cell Line ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Fetal Blood ; cytology ; Fibroblasts ; cytology ; physiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology

The regulation of vascular and trabecular smooth muscle relaxation or contraction in the penis, that is, the physiology of corporal smooth muscle tone, determines penile erection or flaccidity. There is considerable evidence that the potassium channel and calcium channel, like many other vascular tissues, are the major modulators of smooth muscle tone in the corpora. Moreover, data on cultured corporal smooth muscle cells and isolated corporal tissue strips have demonstrated that the neurotransmitters participating in erection modulate corporal smooth muscle tone largely through their effects on ionic channels and transmembrane ionic flux.
Cell Line ; Humans ; Ion Channels ; physiology ; Male ; Muscle Contraction ; physiology ; Muscle, Smooth ; cytology ; physiology ; Penis ; cytology ; physiology ; Potassium Channels ; physiology ; Sodium Channels ; physiology

Animals ; Cell Differentiation ; physiology ; Cell Hypoxia ; Cell Line ; Mice ; Myoblasts ; cytology ; Transcription Factors ; metabolism

With Jingkang No.5 (PiA), calli of the PiA induced for 10-15 days were transferred into amino acid liquid culture medium, to establish excellent rice suspension cell lines successfully in a relative short time. The growth characteristics and differentiation conditions of suspension cells were measured at different phases. Results revealed that the optimal subculture time was 7-10 days, and the cells cultured for 30-120 days had the best differentiation ability (57.1%) and regeneration ability (20%). This study is promising in further using the suspension cell for genetic transformation and protoplasm isolation.
Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Line ; Culture Media ; Oryza ; cytology

Free-living Naegleria fowleri leads to a fatal infection known as primary amebic meningoencephalitis in humans. Previously, the target cell death could be induced by phagocytic activity of N. fowleri as a contact-dependent mechanism. However, in this study we investigated the target cell death under a non-contact system using a tissue-culture insert. The human microglial cells, U87MG cells, co-cultured with N. fowleri trophozoites for 30 min in a non-contact system showed morphological changes such as the cell membrane destruction and a reduction in the number. By fluorescence-activated cell sorter (FACS) analysis, U87MG cells co-cultured with N. fowleri trophozoites in a non-contact system showed a significant increasse of apoptotic cells (16%) in comparison with that of the control or N. fowleri lysate. When U87MG cells were co-cultured with N. fowleri trophozoites in a non-contact system for 30 min, 2 hr, and 4 hr, the cytotoxicity of amebae against target cells was 40.5, 44.2, and 45.6%, respectively. By contrast, the cytotoxicity of non-pathogenic N. gruberi trophozoites was 10.2, 12.4, and 13.2%, respectively. These results suggest that the molecules released from N. fowleri in a contact-independent manner as well as phagocytosis in a contact-dependent manner may induce the host cell death.
Animals ; Apoptosis ; Cell Line ; Humans ; Microglia/*cytology/*parasitology ; Naegleria fowleri/*physiology ; Phagocytosis/physiology

AIMTo explore the effects of tRNA on the growth of mammalian cells.

METHODSL929, NIH3T3, MCF-7 and PC12 cells were seeded in 96 well culture plate individually, and incubated at 37 degrees C in 5% CO2 for 4 h, the tRNAs from different species were added to the culture media individually. After certain time of incubation, the viability of the cells was evaluated by the MTT methods. Sub-confluent L929 cells were incubated with 200 microg/ml ytRNA for different times, then the cells were pooled and analyzed with flow cytometry assay.

RESULTStRNA specifically inhibited the growth of L929 cells in a dose-dependent manner. The sizes of tRNA-treated cells showed larger sizes and longer processes than those of untreated cells. Flow cytometric analysis further showed that most of tRNA-treated cells were arrested in S phase of the cell cycle.

CONCLUSIONThe cell growth inhibitory effects of tRNAs were caused mainly by their degraded fragments. The results suggested that tRNA or its degraded fragments might play important roles in regulation of cell proliferation.

Animals ; Cell Cycle Checkpoints ; physiology ; Cell Line ; Cell Proliferation ; Fibroblasts ; cytology ; Flow Cytometry ; Mice ; RNA, Transfer ; physiology

The vibrational spectral differences of normal and lung cancer cells were studied for the development of effective cancer cell screening by means of attenuated total reflection infrared spectroscopy. The phosphate monoester symmetric stretching nus(PO3(2-)) band intensity at ~970 cm-1 and the phosphodiester symmetric stretching nus(PO2-) band intensity at ~1,085 cm-1 in nucleic acids and phospholipids appeared to be significantly strengthened in lung cancer cells with respect to the other vibrational bands compared to normal cells. This finding suggests that more extensive phosphorylation occur in cancer cells. These results demonstrate that lung cancer cells may be prescreened using infrared spectroscopy tools.
*Carcinoma ; Cell Line, Tumor ; Epithelial Cells/*physiology ; Humans ; *Lung Neoplasms ; Respiratory Mucosa/*cytology ; *Spectrophotometry, Infrared

Human embryonic stem (hES) cells are considered to be a valuable resource for research in regenerative medicine, drug screening, and developmental studies. However, hES cells are usually established and maintained on mouse embryonic fibroblast feeder layers, and the risk of animal origin contamination from feeder layer generally excludes the clinical use of these hES cells. The main emphasis over the last several years has been in finding defined serum-and feeder layer-free system for derivation and culture of hES cells to enable the clinical use of hES cell for cell transplantation.
Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Division ; physiology ; Cell Line ; Coculture Techniques ; Culture Media, Serum-Free ; Embryo Culture Techniques ; Embryo, Mammalian ; cytology ; Embryonic Stem Cells ; physiology ; Humans ; Pluripotent Stem Cells ; cytology ; Stem Cells ; physiology ; Transcription Factors ; physiology ; Transplants