24-Well Plate Lab

Cell culture was a very important area in which HeLa cells were used. Because of HeLa cells, they were able to have large quantities of cancer cells with which to conduct experiments. The cells were given to scientists and researchers who passed them on to others who gave them to even more scientists and researchers. The cells were injected with different drugs in the hopes of finding a cure for cancer and other diseases. HeLa cells did not grow like normal cells but were able to divide an unlimited number of times and provide unlimited amounts of cells for research. During our first semester we learned about cell division and how cells could only divide a certain number of times. HeLa cells were cancer cells taken straight from a cancer tumor and they defied this basic principal of cells division.

In which petri dish did you observe the highest hatching viability? Did the results support your prediction in Pre-Laboratory Question #2?

diH20 acted as a negative control in the experiment, all cells, both in intrafollicular zone and in interfollicular zone stained purple. Cells stained brown in the negative control indicate non-specific staining.

Coat a T75-cm2 culture flasks with 0.2% gelatin to provide better adhesion surface for mEFs.

PC3 (8×104  cells/well) and LNCaP (3×105  cells/well) cells were seeded in 6-well plates in 1.5  mL of complete growth media. Cells were treated with 2.5 μM of free sorafenib or an equivalent sorafenib concentration of SMA-Sor, 3 μM of free nilotinib or an equivalent nilotinib concentration of SMA-Nilo, DMSO or SMA for 48  h. Cell cycle distribution was assessed using propidium iodide staining, as previously described {Somers-Edgar, 2011 #82}. Samples were analysed using a FACScalibur flow cytometer (BD Biosciences, San Jose, CA, USA) and the proportion of cells in each of G0/G1-, S- and G2/M-phases were determined using CellQuest Pro software (BD Biosciences, San Jose, CA, USA).

Transfection of pCas9-GFP with HEK293 cells Introduction Transfection is the insertion of plasmid DNA into eukaryotic cells. Transfection is used to look at the expression of a gene inserted in a cell. For this laboratory, the eukaryotic cells used were HEK293 cells. 293 cells are derived from human embryonic kidney cells.

Introduction Bacteria growth on the petri dishes can be compared to bacteria growth on food. Bacteria grows on food and takes its nutrients from the food, in the same way that the bacteria in the petri dishes gets its nutrients from the nutrient agar. Also bacteria on food can spread very quickly over the food, just as it can in the petri dish. The microorganisms that contaminate food can be good and bad bacteria, as it can be in the petri dish, not all bacteria is harmful. The same common types of bacteria, like molds and yeasts, could be found in both the petri dishes and on spoiled food items.

Human embryonic stem cells (hESCs) are pluripotent and are obtained from the inner mass of a 4-5 day old human blastocyst that consists of approximately 100 cells (“Stem cell research,” 2009).

BMDMs prepared from C57BL/6 mice were re-plated in a non-tissue culture treated 24-well plate at a density of 5 x 105 cells per well on day 6 of differentiation. Twenty-four hours later, the cells were treated with medium only (control), or 10 µg/ml of GP-FITC. After 30 min of incubation at 37°C, the reaction was stopped by washing with ice cold 1x PBS. The adherent cells were detached by incubation with Cellstripper solution

The antibody base method helps detect a specific protein in the sample being studied. IHC helps the researcher visualize the distribution and localization of certain cellular components in the cell. In the experiment, the tissue sections were thawed out for two hours and rehydrated in PBS for five minutes. Triton X-100 was then added to the PBS to help increase the permeability of the cell membrane. The day after, the tissue sections were washed with the PBS, and the sections were then incubated two hours with the secondary antibodies and the nucleic acid staining. The primary antibodies that were used are anti-connexin-43 mouse monoclonal, anti-α-smooth muscle actin mouse monoclonal, and an anti-α-actinin mouse monoclonal. The secondary antibodies that were used were Alexa-fluor antibodies (den Haan et al.,

The recognition of a well-defined ANA staining pattern using the HEp-2 cell substrate may be helpful in

The Kirby-Bauer disk diffusion technique could be used to support this hypothesis for the reason that it will study this frog skin’s impact as an antibiotic against bacteria in stopping it from growing. This can easily be recognized in the disk through a circular area or zone of inhibition seen around the wafers where the bacteria haven’t

This article investigated various effects of Simulated Microgravity (SMG) in mouse embryonic stem cells. The study was conducted by Yulan Wang, Lili An, Yuanda Jiang and Haiying Hang. Funding was provided by grants from the Knowledge Innovation Program of Chinese Academy of Sciences. This study is a follow-up study on previous ones that have been done on adult and differentiated stem cells, but this is the first to be done on embryonic stem cells. Broadly, the two cells types in mammals consist of adult and embryonic stem cells. Adult stem cells are useful in tissue renewal and regeneration after an injury. They conducted this experiment on the embryonic stem cells of mice. Many problems such as skeletal muscle atrophy, cardiovascular problems, immune system dysregulation and alterations of sleep and circadian rhythms are caused by effects of microgravity (MG) at a cellular level (Wang, et al. 2011). The reason microgravity was used is because it has been recognized as a major environmental factor. Because microgravity can only be achieved by entering space, a 3-D clinostat was used to perform the simulation. Many cells types, from bacteria to mammalian cells have been proven to be sensitive to the MG environment, based on past studies. The effects that were monitored included cell cycle distribution, cell differentiation, cell adhesion, apoptosis, genomic integrity and DNA damage repair. The cells cultured under

Cell Culture Contamination is a major problem in the field of scientific research as it obstructs the progression of science greatly. It is the one of the most common problem every researchers have encountered at least once in their lifetime. Cell culture contamination is the very simple process of cell cultures becoming infected with foreign unwanted substances. And these foreign invading substances varies from shapes and sizes to their molecular properties due to the fact that they do not come from a single family of contaminants. Cell cultures often face contamination from chemicals used in laboratories. They however also face contamination from living microorganisms such as bacteria, fungi, mold, etc found in the air and from other cell

The BM-MSCs were cultured in T25 flask culture (Orange Scientific, Braine-l 'Alleud, Belgium) with DMEM medium and incubated for reaching to 60-70% confluency. In the next step, DMEM medium was replaced with RPMI-1640 medium without FBS and was incubated for 72 hours (hrs). After incubation; medium was collected, cells and debris were removed by centrifugation. Supernatant was concentrated by 10 kDa MW cut-off ultrafiltration