scholarly journals Preparation and characterization of nano chitosan for hGM-CSF release

2015 ◽  
Vol 18 (2) ◽  
pp. 64-73
Author(s):  
Truong Tat Dang ◽  
Thuan Cong Nguyen ◽  
Hieu Van Tran
Keyword(s):  
Cell Line ◽  
Average Diameter ◽  
Cell Types ◽  
High Concentration ◽  
Dose Requirement ◽  
Chitosan Beads ◽  
Release Capacity ◽  
Antigen Presenting

hGM-CSF (human granulocytemacrophage colony stimulating factor) is a cytokine secreted by many cell types. Its characters are suitable for vaccine adjuvant such as ability to stimulate survival, differentiation and enhancement the functions of antigen-presenting cells. This cytokine is also a chemoattractant for monocytes and neutrophils to the infected sites, stimulates the expression of several cytokines like IL-1, IL-6, TNF, which are essential for B and T lymphocyte differentiation. However, hGM-CSF has some drawbacks for being an adjuvant candidate due to its easy degradation, toxicity at high concentration and low-dose requirement for therapeutic effect. Drugs delivery system using chitosan can overcome these disadvantages of hGM-CSF. In this present study, chitosan particles were prepared and evaluated the absorption and release of human hGM-CSF. Firstly, the activity of hGM CSF was evaluated by proliferation bioassay using TF-1 cell line. Afterward, chitosan particles were prepared by ionic gelation method and were examined for its toxicity on TF‑1 cell line. After protein absorbance onto chitosan particles, the release capacity and in vitro protection of chitosan for hGM-CSF were assessed. The result showed that hGM-CSF had an ED50 value of 106 pg/mL. The synthesized chitosan particles had an average diameter of 24.5 nm and were nontoxic. Based on the results of SDS-PAGE and Bradford, the adsorption efficiency of hGM‑CSF onto chitosan particles reached 99 % and chitosan has the ability to release hGM-CSF and protects it from hydrolysis of trypsin. In conclusion, the synthesized chitosan beads absorbed and released hGMCSF with its activity remained. This result provides the evidence for further in vivo researches.

scholarly journals Nonmyristoylated Abl proteins transform a factor-dependent hematopoietic cell line.

1992 ◽  
Vol 12 (4) ◽  
pp. 1864-1871 ◽  
Author(s):  
G Q Daley ◽  
R A Van Etten ◽  
P K Jackson ◽  
A Bernards ◽  
D Baltimore
Keyword(s):  
Plasma Membrane ◽  
Cell Line ◽  
Hematopoietic Cells ◽  
Cell Types ◽  
Hematopoietic Cell ◽  
Myelogenous Leukemia ◽  
3T3 Fibroblasts ◽  
Nih 3T3

N-terminal myristoylation can promote the association of proteins with the plasma membrane, a property that is required for oncogenic variants of Src and Abl to transform fibroblastic cell types. The P210bcr/abl protein of chronic myelogenous leukemia cells is not myristoylated and does not stably transform NIH 3T3 fibroblasts; however, it will transform lymphoid and myeloid cell types in vitro and in vivo, suggesting that myristoylation is not required for Abl variants to transform hematopoietic cells. To test this hypothesis, we introduced point mutations that disrupt myristoylation into two activated Abl proteins, v-Abl and a deletion mutant of c-Abl (delta XB), and examined their ability to transform an interleukin-3-dependent lymphoblastoid cell line, Ba/F3. Neither of the nonmyristoylated Abl proteins transformed NIH 3T3 fibroblasts, but like P210bcr/abl, both were capable of transforming the Ba/F3 cells to factor independence and tumorigenicity. Nonmyristoylated Abl variants did not associate with the plasma membrane in the transformed Ba/F3 cells. These results demonstrate that Abl proteins can transform hematopoietic cells in the absence of membrane association and suggest that distinct functions of Abl are required for transformation of fibroblast and hematopoietic cell types.

VARYING RESPONSE TO LUTEINIZING HORMONE OF TWO LUTEAL CELL TYPES ISOLATED FROM BOVINE CORPUS LUTEUM

1979 ◽  
Vol 83 (3) ◽  
pp. 303-NP ◽  
Author(s):  
JOCELYNE URSELY ◽  
PIERRE LEYMARIE
Keyword(s):  
Corpus Luteum ◽  
In Vitro Study ◽  
Average Diameter ◽  
Cell Types ◽  
Enzymatic Digestion ◽  
Cell Suspensions ◽  
Small Cells ◽  
Luteal Cell

Luteal cell suspensions obtained by enzymatic digestion of pregnant cow corpus luteum were found to be heterogenous and mainly made up of two types of cells of different sizes. The large cells (37 μm, average diameter) could be separated from the small ones (18 μm, average diameter) by sedimentation at unit gravity in a gradient of Ficoll–bovine serum albumin. A comparative in-vitro study of the synthesis of progesterone by the two types of cells indicated striking differences between them. The average content and the synthesis of progesterone in the absence and presence of a saturating dose of bovine LH after incubation for 2 h were 0·07, 0·12 and 6·9 pg/cell for the small cells and 0·65, 2 and 10 pg/cell for the large ones. Moreover, the sensitivity to low concentrations of LH was 100 to 1000 times higher for the small cells than for the large ones. oestradiol-17β at concentrations ranging from 5 × 10−10 to 5 × 10−4 mol/l exerted a dose–dependent inhibition on the stimulation of LH in both cell types. These results suggest a possible involvement of both cell types in the synthesis of progesterone in vivo with a greater contribution by the small cells to stimulation induced by LH. Moreover, it appears that small cell suspensions could be a useful model system for in-vitro studies of the control of the synthesis of progesterone in cow corpus luteum.

Identification of toxic mineral dusts using mammalian cells

1980 ◽  
Vol 71 (3) ◽  
pp. 181-184 ◽  
Author(s):  
R. Davies ◽  
M. Chamberlain ◽  
R. C. Brown ◽  
D. M. Griffiths
Keyword(s):  
Cell Line ◽  
Mammalian Cells ◽  
Chinese Hamster ◽  
Cell Types ◽  
Peritoneal Macrophages ◽  
Alveolar Type ◽  
Mineral Dusts ◽  
Potential Pathogenicity

ABSTRACTCell culture systems have been developed to assess the potential pathogenicity of mineral dusts. The in vitro cytotoxicities of a variety of dusts towards mouse peritoneal macrophages, Chinese hamster lung cells (V79 cell line) and human alveolar type II cells (A549 cell line) were investigated.Non-pathogenic dusts were found to be inert in vitro. Fibrogenic non-fibrous dusts such as silica were only cytotoxic towards macrophages. Fibrous dusts which are both fibrogenic and carcinogenic in vivo are cytotoxic towards all three cell types, their cytotoxicity being dependent on fibre size. The size range important for the observed biological effect is longer than about 8 μm and thinner than about 1·5 μm.

scholarly journals NIK-dependent RelB Activation Defines a Unique Signaling Pathway for the Development of Vα14i NKT Cells

2003 ◽  
Vol 197 (12) ◽  
pp. 1623-1633 ◽  
Author(s):  
Dirk Elewaut ◽  
Raziya B. Shaikh ◽  
Kirsten J. L. Hammond ◽  
Hilde De Winter ◽  
Andrew J. Leishman ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Cell Types ◽  
Nkt Cells ◽  
Compound Heterozygous ◽  
Nkt Cell ◽  
Surface Receptors ◽  
Development In Vitro ◽  
Antigen Presenting

A defect in RelB, a member of the Rel/nuclear factor (NF)-κB family of transcription factors, affects antigen presenting cells and the formation of lymphoid organs, but its role in T lymphocyte differentiation is not well characterized. Here, we show that RelB deficiency in mice leads to a selective decrease of NKT cells. RelB must be expressed in an irradiation-resistant host cell that can be CD1d negative, indicating that the RelB expressing cell does not contribute directly to the positive selection of CD1d-dependent NKT cells. Like RelB-deficient mice, aly/aly mice with a mutation for the NF-κB–inducing kinase (NIK), have reduced NKT cell numbers. An analysis of NK1.1 and CD44 expression on NKT cells in the thymus of aly/aly mice reveals a late block in development. In vitro, we show that NIK is necessary for RelB activation upon triggering of surface receptors. This link between NIK and RelB was further demonstrated in vivo by analyzing RelB+/− × aly/+ compound heterozygous mice. After stimulation with α-GalCer, an antigen recognized by NKT cells, these compound heterozygotes had reduced responses compared with either RelB+/− or aly/+ mice. These data illustrate the complex interplay between hemopoietic and nonhemopoietic cell types for the development of NKT cells, and they demonstrate the unique requirement of NKT cells for a signaling pathway mediated by NIK activation of RelB in a thymic stromal cell.

scholarly journals Nonmyristoylated Abl proteins transform a factor-dependent hematopoietic cell line

1992 ◽  
Vol 12 (4) ◽  
pp. 1864-1871
Author(s):  
G Q Daley ◽  
R A Van Etten ◽  
P K Jackson ◽  
A Bernards ◽  
D Baltimore
Keyword(s):  
Plasma Membrane ◽  
Cell Line ◽  
Hematopoietic Cells ◽  
Cell Types ◽  
Hematopoietic Cell ◽  
Myelogenous Leukemia ◽  
3T3 Fibroblasts ◽  
Nih 3T3

N-terminal myristoylation can promote the association of proteins with the plasma membrane, a property that is required for oncogenic variants of Src and Abl to transform fibroblastic cell types. The P210bcr/abl protein of chronic myelogenous leukemia cells is not myristoylated and does not stably transform NIH 3T3 fibroblasts; however, it will transform lymphoid and myeloid cell types in vitro and in vivo, suggesting that myristoylation is not required for Abl variants to transform hematopoietic cells. To test this hypothesis, we introduced point mutations that disrupt myristoylation into two activated Abl proteins, v-Abl and a deletion mutant of c-Abl (delta XB), and examined their ability to transform an interleukin-3-dependent lymphoblastoid cell line, Ba/F3. Neither of the nonmyristoylated Abl proteins transformed NIH 3T3 fibroblasts, but like P210bcr/abl, both were capable of transforming the Ba/F3 cells to factor independence and tumorigenicity. Nonmyristoylated Abl variants did not associate with the plasma membrane in the transformed Ba/F3 cells. These results demonstrate that Abl proteins can transform hematopoietic cells in the absence of membrane association and suggest that distinct functions of Abl are required for transformation of fibroblast and hematopoietic cell types.

Ultrastructural Study of a differentiating human colon carcinoma cell line

1990 ◽  
Vol 48 (3) ◽  
pp. 208-209
Author(s):  
Sylvie Polak-Charcon ◽  
Mehrdad Hekmati ◽  
Yehuda Ben Shaul
Keyword(s):  
Cell Line ◽  
Morphological Changes ◽  
Secretory Granules ◽  
Human Colon ◽  
Cell Types ◽  
Culture Conditions ◽  
Morphological Evidence ◽  
Human Colon Carcinoma Cell ◽  
Colon Cell

The epithelium of normal human colon mucosa “in vivo” exhibits a gradual pattern of differentiation as undifferentiated stem cells from the base of the crypt of “lieberkuhn” rapidly divide, differentiate and migrate toward the free surface. The major differentiated cell type of the intestine observed are: absorptive cells displaying brush border, goblet cells containing mucous granules, Paneth and endocrine cells containing dense secretory granules. These different cell types are also found in the intestine of the 13-14 week old embryo.We present here morphological evidence showing that HT29, an adenocarcinoma of the human colon cell line, can differentiate into various cell types by changing the growth and culture conditions and mimic morphological changes found during development of the intestine in the human embryo.HT29 cells grown in tissue-culture dishes in DMEM and 10% FCS form at late confluence a multilayer of morphologically undifferentiated cell culture covered with irregular microvilli, and devoid of tight junctions (Figs 1-3).

Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

1990 ◽  
Vol 48 (3) ◽  
pp. 914-915
Author(s):  
D.J.P. Ferguson ◽  
A.R. Berendt ◽  
J. Tansey ◽  
K. Marsh ◽  
C.I. Newbold
Keyword(s):  
Plasmodium Falciparum ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Umbilical Vein ◽  
Cell Types ◽  
Amelanotic Melanoma ◽  
Cytoplasmic Process ◽  
Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

Epithelial Organotypic Cultures: A Viable Model to Address Mechanisms of Carcinogenesis by Epitheliotropic Viruses

2018 ◽  
Vol 18 (4) ◽  
pp. 246-255 ◽  
Author(s):  
Lara Termini ◽  
Enrique Boccardo
Keyword(s):  
Three Dimensional ◽  
Cell Types ◽  
Experimental Models ◽  
Biological Research ◽  
Specific Gene ◽  
Specific Cell ◽  
Organotypic Cultures ◽  
Skin Physiology

In vitro culture of primary or established cell lines is one of the leading techniques in many areas of basic biological research. The use of pure or highly enriched cultures of specific cell types obtained from different tissues and genetics backgrounds has greatly contributed to our current understanding of normal and pathological cellular processes. Cells in culture are easily propagated generating an almost endless source of material for experimentation. Besides, they can be manipulated to achieve gene silencing, gene overexpression and genome editing turning possible the dissection of specific gene functions and signaling pathways. However, monolayer and suspension cultures of cells do not reproduce the cell type diversity, cell-cell contacts, cell-matrix interactions and differentiation pathways typical of the three-dimensional environment of tissues and organs from where they were originated. Therefore, different experimental animal models have been developed and applied to address these and other complex issues in vivo. However, these systems are costly and time consuming. Most importantly the use of animals in scientific research poses moral and ethical concerns facing a steadily increasing opposition from different sectors of the society. Therefore, there is an urgent need for the development of alternative in vitro experimental models that accurately reproduce the events observed in vivo to reduce the use of animals. Organotypic cultures combine the flexibility of traditional culture systems with the possibility of culturing different cell types in a 3D environment that reproduces both the structure and the physiology of the parental organ. Here we present a summarized description of the use of epithelial organotypic for the study of skin physiology, human papillomavirus biology and associated tumorigenesis.

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