Isolation and culture of bovine oviductal epithelial cells for use in the anatomy and physiology laboratory and undergraduate research

2006 ◽  
Vol 30 (4) ◽  
pp. 237-241 ◽  
Author(s):  
Amy L. Way
Keyword(s):  
Cell Culture ◽  
Epithelial Cells ◽  
Culture Medium ◽  
Undergraduate Research ◽  
Teaching Laboratory ◽  
Anatomy And Physiology ◽  
Undergraduate Research Program ◽  
Physiology Laboratory ◽  
Experimental Use ◽  
Oviductal Cell

This article presents methods for the isolation and culture of epithelial cells from the bovine oviduct for use in both research and the teaching laboratory and provides examples of ways that an oviductal cell culture can be incorporated into an undergraduate research program. Cow reproductive tracts are readily available from area butchers, and the procedure for isolation of the epithelium is simple and inexpensive. The cells can be observed immediately after isolation or can be cultured for up to 72 h under simple conditions for observation over several days. For experimental use, epithelial cells are cultured in standard cell culture medium, where they continue to divide and actively secrete substances into the medium. The ease with which the tissue can be collected and cells isolated makes the oviductal epithelium ideal for use in both the teaching laboratory and research projects in which undergraduates serve as investigators.

Relationships between Undergraduate Research and the Advanced Undergraduate Teaching Laboratory

2015 ◽  
Vol 1762 ◽  
Author(s):  
Colin Inglefield
Keyword(s):  
Research Program ◽  
Undergraduate Education ◽  
Materials Science ◽  
Undergraduate Research ◽  
Undergraduate Teaching ◽  
Complementary Relationship ◽  
Teaching Laboratory ◽  
Undergraduate Research Program ◽  
Research Techniques ◽  
A Priority

ABSTRACTI will present a model for a complementary relationship between an undergraduate research program and the undergraduate teaching laboratory in materials science. One clear example is using the teaching laboratory to prepare students to do more independent undergraduate research by emphasizing appropriate skills and knowledge, but there are several others. Undergraduate researchers can work with faculty to develop novel experiments and apparatus that can be used in the teaching laboratory. Undergraduates can gain working knowledge of common research techniques and equipment within their program. The ideas should be applicable to any institution placing a priority on undergraduate research and undergraduate education.

124 CRYOPRESERVED BOVINE OVIDUCTAL EPITHELIAL CELLS SURVIVAL, GROWTH, AND ABILITY TO SUPPORT EARLY EMBRYO DEVELOPMENT

2015 ◽  
Vol 27 (1) ◽  
pp. 154
Author(s):  
E. Corbin ◽  
A. Cordova ◽  
J. Grosbois ◽  
P. Mermillod
Keyword(s):  
Cell Culture ◽  
Epithelial Cells ◽  
Embryo Development ◽  
Culture Medium ◽  
Early Embryo ◽  
Cell Culture Medium ◽  
Cleavage Rate ◽  
Early Embryo Development ◽  
Blastocyst Rate

Previous experiments demonstrated that co-culture of bovine embryos with bovine oviducal epithelial cells (BOEC) improved blastocyst rate and quality (Cordova et al. 2014). However, the use of primary cell support for improving embryo development in vitro may introduce a higher variability of the results between different BOEC batches used, as well as sanitary risks. The use of well-controlled large batches of frozen BOEC may help to solve these problems. Therefore, the aim of the present study was to characterise the survival and functionality of frozen-thawed BOEC. Bovine oviducts attached to ovaries showing recent ovulation were collected at a local slaughterhouse during 4 replicates (3 oviducts per replicate). Epithelial cells were expelled by gentle squeezing and washed 3 times. Half of the cell pellet was diluted 100-fold in culture medium (TCM199 + 10% FCS) for culture of fresh cells. The other half was diluted 10-fold in cell freezing medium (TCM199 + 20% FCS + 10% dimethyl sulfoxide), allowed to equilibrate in this medium for 10 min, and frozen at –80°C in a container filled with isopropyl alcohol. After 4 h, the tubes were transferred into LN for at least 1 h. The tubes were then thawed (5 min in 37°C water bath), diluted 1 : 1 in cell culture medium, and centrifuged for 10 min at 100 × g. The pellet was then diluted 100× in cell culture medium. Fresh or frozen-thawed cells were seeded in 4-well NUNC plates for 7 days at 38.8°C in a humidified atmosphere with 5% CO2 in air. The medium was renewed every 48 h, and the viability of cells was assessed by calcein-AM and ethidium homodimer labelling. After 7 days of culture, the medium was replaced by SOF medium + 5% FCS, and bovine in vitro-produced zygotes were added the day after and co-cultured for 8 days at 38.8°C in a humidified atmosphere with 5% CO2 in air to evaluate embryo development. Half of the medium was renewed every 48 h. Frozen-thawed cells showed the same viability than fresh ones at Days 0 and 7 of culture and reached confluence at the same time (Day 7). Development results are shown in Table 1. Frozen and fresh cells support early embryo development at the same rate. In conclusion, the present study showed that BOEC frozen on the day of collection are equivalent to fresh BOEC in regards to their survival and proliferation and their ability to support early embryo development. At collection, the cells may face stresses that are just as considerable as freezing/thawing (temperature shock, scrapping, change of environment). This may explain why they are not affected by freezing than at collection. The differentiation status of these cells is now under analysis by immunocytochemistry. Table 1.Cleavage rate and blastocyst rate in 3 different types of culture systems

Fibronectin and its alpha 5 beta 1-integrin receptor are involved in the wound-repair process of airway epithelium

1996 ◽  
Vol 271 (5) ◽  
pp. L726-L733 ◽  
Author(s):  
A. L. Herard ◽  
D. Pierrot ◽  
J. Hinnrasky ◽  
H. Kaplan ◽  
D. Sheppard ◽  
...  
Keyword(s):  
Cell Culture ◽  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Airway Epithelium ◽  
Wound Repair ◽  
Culture Medium ◽  
Type I ◽  
Integrin Subunit ◽  
Wound Area ◽  
Beta 1 Integrin

The cell migration that occurs during wound repair is dependent on modifications of the cell-matrix interaction in which extracellular matrix proteins and their receptors, the integrins, are involved. To study the interactions between airway epithelial cells and the extracellular matrix during the process of wound repair, we developed an in vitro wound model of human epithelial cells. Surface epithelial cells were dissociated from human nasal polyps and cultured on a type I collagen matrix. At confluency, a wound was made by the addition of 2 microliters of NaOH (1 N) to the cell culture. After the cell culture was washed, the wound area was recorded every 12 h for 96 h by a videomicroscopic technique. We calculated the wound-repair index that represents the decrease in the wound area per hour. Using immunofluorescence techniques, we first examined the localization, during wound repair, of fibronectin and of the beta 1-, alpha v-, alpha 2-, alpha 3-, and alpha 5-integrin subunits. Secondly, we carried out a series of wound-repair blocking experiments with the use of anti-integrin or anti-fibronectin antibodies diluted in the culture medium. We observed that fibronectin and the alpha 5- integrin subunit were exclusively expressed by the migratory cells in the wounded area. No difference in the localization of the alpha v-, alpha 2-, and alpha 3-integrin subunits was observed between the nonrepairing and repairing cells. The blocking experiments showed a significant decrease in the wound-repair index in the presence of either the anti-beta 1, -alpha 3, alpha 5, or the anti-fibronectin antibodies. Furthermore, the addition of fibronectin to the culture medium induced a significant increase in the wound repair index. These results suggest that fibronectin and the corresponding alpha 5 beta 1-integrin play an important role in the process of airway epithelium wound repair.

3-iodothyronamine (T1AM) is taken up and rapidly metabolized in cell culture medium only at low concentration

2020 ◽  
Author(s):  
Federica Saponaro ◽  
Marco Borsò ◽  
Sara Verlotta ◽  
Lavinia Bandini ◽  
Alessandro Saba ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Low Concentration

Effect of Atmospheric-Pressure Helium Plasma Jet on Cell Culture Medium

2013 ◽  
Vol 133 (5) ◽  
pp. 278-285
Author(s):  
Norimitsu Takamura ◽  
Douyan Wang ◽  
Takao Satoh ◽  
Takao Namihira ◽  
Hisato Saitoh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Atmospheric Pressure ◽  
Culture Medium ◽  
Plasma Jet ◽  
Helium Plasma ◽  
Cell Culture Medium

scholarly journals Undergraduate Research Program to Recycle Composite Waste

2021 ◽  
Vol 11 (7) ◽  
pp. 354
Author(s):  
Waleed Ahmed ◽  
Essam Zaneldin ◽  
Amged Al Hassan
Keyword(s):  
Mechanical Properties ◽  
Compressive Strength ◽  
Carbon Fiber ◽  
Undergraduate Students ◽  
Research Program ◽  
Modulus Of Elasticity ◽  
Undergraduate Research ◽  
Undergraduate Research Program ◽  
Structural Applications ◽  
Cfrp Composite

With the rapid growth in the manufacturing industry and increased urbanization, higher amounts of composite material waste are being produced, causing severe threats to the environment. These environmental concerns, coupled with the fact that undergraduate students typically have minimal experience in research, have initiated the need at the UAE University to promote research among undergraduate students, leading to the development of a summer undergraduate research program. In this study, a recycling methodology is presented to test lab-fabricated Carbon-Fiber-Reinforced Polymer (CFRP) for potential applications in industrial composite waste. The work was conducted by two groups of undergraduate students at the UAE University. The methodology involved the chemical dissolution of the composite waste, followed by compression molding and adequate heat treatment for rapid curing of CFRP. Subsequently, the CFRP samples were divided into three groups based on their geometrical distinctions. The mechanical properties (i.e., modulus of elasticity and compressive strength) were determined through material testing, and the results were then compared with steel for prompt reference. The results revealed that the values of mechanical properties range from 2 to 4.3 GPa for the modulus of elasticity and from 203.7 to 301.5 MPa for the compressive strength. These values are considered competitive and optimal, and as such, carbon fiber waste can be used as an alternate material for various structural applications. The inconsistencies in the values are due to discrepancies in the procedure as a result of the lack of specialized equipment for handling CFRP waste material. The study concluded that the properties of CFRP composite prepreg scrap tend to be reusable instead of disposable. Despite the meager experimental discrepancies, test values and mechanical properties indicate that CFRP composite can be successfully used as a material for nonstructural applications.

scholarly journals Defined MSC exosome with high yield and purity to improve regenerative activity

2021 ◽  
Vol 12 ◽  
pp. 204173142110086
Author(s):  
Jun Yong Kim ◽  
Won-Kyu Rhim ◽  
Yong-In Yoo ◽  
Da-Seul Kim ◽  
Kyoung-Won Ko ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Density Lipoprotein ◽  
High Yield ◽  
Human Umbilical Cord ◽  
Cell Culture Medium ◽  
Human Coronary Artery ◽  
Isolation Methods ◽  
Regenerative Activity ◽  
Yield And Purity

Exosomes derived from mesenchymal stem cells (MSCs) have been studied as vital components of regenerative medicine. Typically, various isolation methods of exosomes from cell culture medium have been developed to increase the isolation yield of exosomes. Moreover, the exosome-depletion process of serum has been considered to result in clinically active and highly purified exosomes from the cell culture medium. Our aim was to compare isolation methods, ultracentrifuge (UC)-based conventional method, and tangential flow filtration (TFF) system-based method for separation with high yield, and the bioactivity of the exosome according to the purity of MSC-derived exosome was determined by the ratio of Fetal bovine serum (FBS)-derived exosome to MSC-derived exosome depending on exosome depletion processes of FBS. The TFF-based isolation yield of exosome derived from human umbilical cord MSC (UCMSC) increased two orders (92.5 times) compared to UC-based isolation method. Moreover, by optimizing the process of depleting FBS-derived exosome, the purity of UCMSC-derived exosome, evaluated using the expression level of MSC exosome surface marker (CD73), was about 15.6 times enhanced and the concentration of low-density lipoprotein-cholesterol (LDL-c), known as impurities resulting from FBS, proved to be negligibly detected. The wound healing and angiogenic effects of highly purified UCMSC-derived exosomes were improved about 23.1% and 71.4%, respectively, with human coronary artery endothelial cells (HCAEC). It suggests that the defined MSC exosome with high yield and purity could increase regenerative activity.

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