How Does The Bone Marrow Mesenchymal Stem Cell Experiment
MATERIAL AND METHODS
1. Materials: 1.1. Experimental animals:
The experiment was carried out on 30 adult healthy female rats weighing 190 20g and 10 young (6-7) weeks –old male albino rats from which bone marrow mesenchymal stem cell (MSCs) were isolated. Animal were purchased from the animal house of the faculty of Medicine Mansoura University. The animals were housed in standard stainless steel cages at room temperature 25-28ﾟC, humidity of (50-60%) and maintained at 12 hrs alternating day and night cycle. The rats were fed standard pellet diet and water ad libitum.
The rats were acclimatized to their housing and feeding for two weeks before the start of the experiment. The local committee approved the design of the experiments and the protocols were carried out according to the guidelines of the National Institutes of Health (NIH). …show more content…
• The tissue culture flasks containing bone marrow and complete culture medium, were incubated in a standard CO2 incubator (SHEL- LAB CO2 Incubator) at 100% humidity,37ºC in 5% CO2 and 95% O2 (by volume) for at least three days before the media changed. (Fig. 36.e.).
• The cultured cells were examined daily by using the inverted microscope (OLYMPUS-CKX41) (Fig. 36.f.) to follow up the growth of the cells and to detect the appearance of any bacterial or fungal infection among the culture cells. • The medium was first changed on the third day of culture and every other 2-3 days. The supernatant that contained the floating non-adherent cells was removed by aspiration using a sterile pipette. The adherent cells were then washed twice with PBS. Finally 25 ml of fresh complete culture medium was added to the tissue culture flasks. The MSCS were distinguished from other bone marrow cells by their tendency to adhere to tissue culture
1. Materials: 1.1. Experimental animals:
The experiment was carried out on 30 adult healthy female rats weighing 190 20g and 10 young (6-7) weeks –old male albino rats from which bone marrow mesenchymal stem cell (MSCs) were isolated. Animal were purchased from the animal house of the faculty of Medicine Mansoura University. The animals were housed in standard stainless steel cages at room temperature 25-28ﾟC, humidity of (50-60%) and maintained at 12 hrs alternating day and night cycle. The rats were fed standard pellet diet and water ad libitum.
The rats were acclimatized to their housing and feeding for two weeks before the start of the experiment. The local committee approved the design of the experiments and the protocols were carried out according to the guidelines of the National Institutes of Health (NIH). …show more content…
• The tissue culture flasks containing bone marrow and complete culture medium, were incubated in a standard CO2 incubator (SHEL- LAB CO2 Incubator) at 100% humidity,37ºC in 5% CO2 and 95% O2 (by volume) for at least three days before the media changed. (Fig. 36.e.).
• The cultured cells were examined daily by using the inverted microscope (OLYMPUS-CKX41) (Fig. 36.f.) to follow up the growth of the cells and to detect the appearance of any bacterial or fungal infection among the culture cells. • The medium was first changed on the third day of culture and every other 2-3 days. The supernatant that contained the floating non-adherent cells was removed by aspiration using a sterile pipette. The adherent cells were then washed twice with PBS. Finally 25 ml of fresh complete culture medium was added to the tissue culture flasks. The MSCS were distinguished from other bone marrow cells by their tendency to adhere to tissue culture