394 DEVELOPMENT OF A RAT ENDOMETRIAL SPHEROID AS A MODEL FOR THE ANALYSIS OF ENDOMETRIAL FUNCTIONS

2007 ◽  
Vol 19 (1) ◽  
pp. 312
Author(s):  
K. Nishimura ◽  
S. Oozono ◽  
H. Urata ◽  
P. He ◽  
T. Soh ◽  
...  
Keyword(s):  
Cell Biology ◽  
In Vitro Model ◽  
Monolayer Culture ◽  
Cell Sheet ◽  
Nuclear Antigen ◽  
Marker Genes ◽  
Positive Staining ◽  
Cell Sheets ◽  
Vitro Model

The endometrium is one of the most complex tissues; it undergoes dynamic changes in response to implantation and pregnancy processes.An in vitro model may provide a tool for clarifying the complex implantation process. However, there is no suitable in vitro model for investigation of endometrial functions. The spheroid has been utilized in cell biology research because it appears to mimic the morphology and physiology of cells in living tissues and organs, which is unlike conventional monolayer culture. Multicellular spheroids composed of normal adult cells may provide a more useful model for the study of the endometrium. The purpose of the present study was to develop a spheroid composed of rat endometrial stromal (RES) cells as an in vitro model for analysis of endometrial functions. The RES were prepared from rat endometrium at Day 5 of pregnancy. Spheroids were generated using salmon aterocollagen (SAC). The cells were plated on 12-well SAC gels (Imoto Suisan Co.) and cultured in DMEM/F12 containing 10% fetal bovine serum. After the cells reached confluence at Day 7 in culture, SAC gels were digested by collagenase to promote the detachment of RES cell sheets. Then the floating cell sheets were transferred to agarose-coated plates and cultured to form spheroids. The cell sheet shrank and became an aggregated cell mass in a few days; it finally formed a round-shaped spheroid. Diameters of the spheroids were about 516.7 � 28.9 �m at Day 5 after detachment from SAC gels. TUNEL examination of cell viability in the spheroid suggested that no cell was apoptotic until 15 days after cell sheet detachment; TUNEL-positive cells appeared at 20 days. Additionally, no positive staining of proliferating cell nuclear antigen (PCNA) was observed in the spheroids in contrast to strong staining in proliferating monolayer cultured cells. The results of gelatin zymography showed that both matrix metalloproteinases (MMP)-2 and-9 were produced in monolayer culture. However, after the detachment of the cell sheet, the production of both MMPs decreased immediately and could not be detected until 15 days after detachment. In vitro decidualization of the spheroids was induced by arachidonic acid (AA) treatment. RT-PCR analysis showed that typical marker genes for the decidualization, desmin and decidual/trophoblast prolactin-related protein (d/tPRP), were expressed in the spheroids after 2 days of AA treatment, but not in the control groups without treatment. The results of the present study indicate that rat endometrial stromal cells are capable of being regenerated as a spheroid using SAC gels in vitro. Importantly, the present spheroid displays an endometrium-mimicking feature in both structural and functional similarities. The present method is simple and convenient, and therefore provides a new insight into the study of endometrial functions and implantation. This research was supported by a grant from the Ministry of Education, Culture, Sport, Science, and Technology of Japan (Kiban-kenkyu C 18580282).

scholarly journals Microencapsulated multicellular tumor spheroids:preparation and use as a novel in vitro model for drug screening

2010 ◽  
Vol 56 (6) ◽  
pp. 674-685 ◽  
Author(s):  
A.M. Tsoy ◽  
D.S. Zaytseva-Zotova ◽  
E.F. Edelweiss ◽  
A. Bartkowiak ◽  
J-L. Goergen ◽  
...  
Keyword(s):  
Drug Screening ◽  
In Vitro Model ◽  
Monolayer Culture ◽  
Breast Adenocarcinoma ◽  
Viable Cells ◽  
Adenocarcinoma Cells ◽  
Mean Size ◽  
Vitro Model ◽  
Mcf 7

In the current study a technique for microencapsulation of human breast adenocarcinoma cells MCF-7 in alginate-chitosan microcapsules is used. Microencapsulation is proposed to generate multicellular tumor spheroids (MTS) based on these cells and to test them further as an in vitro model for anti-tumor drug screening. Cytotoxicity of methotrexate (MTX) was studied on the obtained MTS. A set of MTS with mean size of 150, 200 and 300 m was prepared in function of a cultivation time. After incubation of MTS in cultivation medium containing MTX at concentrations of 1, 2, 10, 50 and 100 nM for 48 hs cell viability was evaluated. MTS were shown to be more resistant to MTX than the monolayer culture, and the resistance to MTX was increased with enhancing a spheroid size. At MTX concentration of 100 nM a number of viable cells in MTS with the size of 300 m was 2.5-fold bigger than that one in monolayer culture. It is suggested that the cells in microencapsulated MTS can better mimic cell behavior in a small size solid tumor than the cells in a monolayer culture. In future microencapsulated MTS can be proposed as a novel in vitro model for anticancer drug screening.

scholarly journals Imaging cell biology in live animals: Ready for prime time

2013 ◽  
Vol 201 (7) ◽  
pp. 969-979 ◽  
Author(s):  
Roberto Weigert ◽  
Natalie Porat-Shliom ◽  
Panomwat Amornphimoltham
Keyword(s):  
Cell Biology ◽  
Cancer Biology ◽  
In Vitro Model ◽  
Time Lapse ◽  
Living Cells ◽  
Model Systems ◽  
Prime Time ◽  
Vitro Model

Time-lapse fluorescence microscopy is one of the main tools used to image subcellular structures in living cells. Yet for decades it has been applied primarily to in vitro model systems. Thanks to the most recent advancements in intravital microscopy, this approach has finally been extended to live rodents. This represents a major breakthrough that will provide unprecedented new opportunities to study mammalian cell biology in vivo and has already provided new insight in the fields of neurobiology, immunology, and cancer biology.

Quantitative Analysis of Unidirectional 2-D Tissue Formation of Endothelial Cells

1992 ◽  
Vol 1 (6) ◽  
pp. 375-381 ◽  
Author(s):  
Satoshi Niu ◽  
Takehisa Matsuda
Keyword(s):  
Endothelial Cells ◽  
Dynamic Process ◽  
Vascular Graft ◽  
In Vitro Model ◽  
Cell Sheet ◽  
Glass Slide ◽  
Tissue Formation ◽  
Culture Chamber ◽  
Vitro Model

We developed a reliable and quantitative method for measuring the dynamic process of unidirectional two-dimensional (2-D) tissue formation of endothelial cells (ECs) in vitro. The culturing of bovine ECs in an assembled culture chamber provided a square monolayered cell sheet with a linear margin when disassembled at the confluency. The cell sheet maintained in culture showed a unidirectional endothelialization in vitro. The cell population-distance histogram, which was determined from the daily observation of tissue, allowed us to determine quantitatively the dynamic process of unidirectional endothelialization in vitro. The endothelialized distance and the endothelializing zone on a glass slide were found to be nearly 500 μm/day and 750 μm in width, respectively. Thus, the method developed here provided information of the 2-D tissue formation process. This model would be useful as an in vitro model which simulates the anastomotic endothelialization of an artificial vascular graft.

scholarly journals Biology of the cardiac myocyte in heart disease

2016 ◽  
Vol 27 (14) ◽  
pp. 2149-2160 ◽  
Author(s):  
Angela K. Peter ◽  
Maureen A. Bjerke ◽  
Leslie A. Leinwand
Keyword(s):  
Cell Biology ◽  
In Vitro Model ◽  
Cardiac Myocyte ◽  
Model Systems ◽  
Human Stem Cells ◽  
Cell Type ◽  
Pathological Hypertrophy ◽  
Vitro Model ◽  
Immortalized Cell

Cardiac hypertrophy is a major risk factor for heart failure, and it has been shown that this increase in size occurs at the level of the cardiac myocyte. Cardiac myocyte model systems have been developed to study this process. Here we focus on cell culture tools, including primary cells, immortalized cell lines, human stem cells, and their morphological and molecular responses to pathological stimuli. For each cell type, we discuss commonly used methods for inducing hypertrophy, markers of pathological hypertrophy, advantages for each model, and disadvantages to using a particular cell type over other in vitro model systems. Where applicable, we discuss how each system is used to model human disease and how these models may be applicable to current drug therapeutic strategies. Finally, we discuss the increasing use of biomaterials to mimic healthy and diseased hearts and how these matrices can contribute to in vitro model systems of cardiac cell biology.

An in vitro model for investigation of vitamin A effects on wound healing

2021 ◽  
pp. 1-6
Author(s):  
Hoda Keshmiri Neghab ◽  
Mohammad Hasan Soheilifar ◽  
Gholamreza Esmaeeli Djavid
Keyword(s):  
Wound Healing ◽  
Endothelial Cell ◽  
Vitamin A ◽  
In Vitro Model ◽  
Cellular Proliferation ◽  
Endothelial Cell Migration ◽  
Normal Fibroblast ◽  
Anti Inflammatory ◽  
Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inflammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inflammation responses.

A new in vitro model for pre-transplantation diagnosis of frozen/thawed human ovarian tissue

2011 ◽  
Vol 71 (05) ◽  
Author(s):  
M Salama ◽  
K Winkler ◽  
KF Murach ◽  
S Hofer ◽  
L Wildt ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Ovarian Tissue ◽  
Human Ovarian Tissue ◽  
Vitro Model
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