Isolation, Characterization and Growth Kinetic Comparison of Bone Marrow and Adipose Tissue Mesenchymal Stem Cells of Guinea Pig

BACKGROUND: Mesenchymal stem cells (MSCs) from different sources have different characteristics. Moreover, MSCs are not isolated and characterized in Guinea pig for animal model of cell therapy. AIM OF THE WORK: was the isolating of bone marrow MSCs (BM-MSCs) and adipose tissue MSCs (AT-MSCs) from Guinea pig and assessing their characteristics. MATERIAL AND METHODS: In this study, bone marrow and adipose tissue were collected from three Guinea pigs and cultured and expanded through eight passages. BM-MSCs and AT-MSCs at passages 2, 5 and 8 were seeded in 24-well plates in triplicate. Cells were counted from each well 1~7 days after seeding to determine population doubling time (PDT) and cell growth curves. Cells of passage 3 were cultured in osteogenic and adipogenic differentiation media. RESULTS: BM-MSCs and AT-MSCs attached to the culture flask and displayed spindle-shaped morphology. Proliferation rate of AT-MSCs in the analyzed passages was more than BM-MSCs. The increase in the PDT of MSCs occurs with the increase in the number of passages. Moreover, after culture of BM-MSCs and AT-MSCs in differentiation media, the cells differentiated toward osteoblasts and adipocytes as verified by Alizarin Red staining and Oil Red O staining, respectively. CONCLUSION: BM-MSCs and AT-MSCs of Guinea pig could be valuable source of multipotent stem cells for use in experimental and preclinical studies in animal models.

Growth Kinetics, characterization, and plasticity of human menstrual blood stem cells

One of the readily available sources of mesenchymal stem cells [MSCs] is menstrual blood-derived stem cells [Men-SCs], which exhibit characteristics similar to other types of MSCs. This study was performed to determine the growth kinetics, plasticity, and characterization of Men-SCs in women. During spring 2014 in the southern Iranian city of Shiraz, menstrual blood [5 mL] was obtained from 10 women on their third day of menstruation in 2 age groups of 30 to 40 and 40 to 50 years old. Ficoll was used to separate the mononuclear cell fraction. After the Men-SCs were cultured, they were subcultured up to passage 4. Growth behavior and population doubling time were evaluated by seeding 5×10[4] cells into 12- and 24-well culture plates, and the colonies were enumerated. The expression of CD44, CD90, and CD34 was evaluated. The osteogenic potential was assessed by alizarin red staining. The Men-SCs were shown to be plastic adherent and spindle-shaped. Regarding the growth curves in the 12-24-well culture plates, it was demonstrated that in the women aged between 30 and 40 years, population doubling time was 55.5 and 62 hours, respectively, while these values in the women aged between 40 and 50 years were 70.4 and 72.4 hours, correspondingly. Positive expression of CD44 and CD90 and negative expression of CD34 were noted. In the osteogenic differentiation medium, the cells differentiated toward osteoblasts. As human Men-SCs are easily collectable without any invasive procedure and are a safe and rapid source of MSCs, they can be a good candidate for stem cell banking and cell transplantation in women

Effects of recombinant human erythropoietin pretreatment on anti HLA antibody titer: a preclinical experience in rats

Erythropoietin [EPO] was first known as a factor for red blood cell proliferation and differentiation. New studies show the effects of EPO on immune system. In this study, the effects of pretreatment with recombinant human erythropoietin [rHuEPO] on the anti-human leukocyte antibody [anti-HLA] titer were determined. Three groups of rats were sensitized with human lymphocytes. Two of the groups were given 20 or 100 IU/Kg rHuEPO after two sensitizations with human lymphocytes. Control group did not receive rHuEPO. Microlymphocytotoxicity method was used to detect anti-HLA antibodies. Treatment with rHuEPO caused a significant decline in anti-HLA antibody titer compared to control group. Also, pretreatment with rHuEPO suppressed antibody response after repeated antigenic stimulation. Such results could be due to the effects of rHuEPO on the number or the activity of the B and the T cells. Moreover, the dose of rHuEPO and the length of treatment might affect anti-HLA antibody titer

Effects of hydroalchoholic extract of anethum graveolens [DILL] on plasma glucose an lipid levels in diabetes induced rats

Type 1 diabetes or insulin dependent diabetes mellitus is an autoimmune disease probably caused by various factors such genetics, environmental stresses, viral infections and diet. The disease is associated with damage to pancreatic insulin-producing beta cells. Many researches have been performed to study the therapeutic effects of herbal medications on diabetes mellitus. The protective effects of some plants such as Cuminum cyminum, Trigonella foenum-graecum, Saliva sclarea, Allium ampeloprasum etc in diabetes induced rats were reported. Dill with the scientific name of Anethum graveolens, is a plant from umbellifera. This family which contains flavonoid compounds. In this research, effects of hydroalchoholic extract of this plant on blood glucose level, lipids and lipoproteins in Alloxan induced diabetic rats was investigated. Fifteen adult male rats, weighing 200-250 gr, were used in three five member groups. Rats in the control group, received physiological serum, as intraperitoneal [IP]. In diabetic control group [DCG], diabetes was induced using Alloxan monohydrate at a dose of 120 mg/kgbw. In addition to diabetes induction just like DCG, the treated diabetic control group [TDCG] rats were dosed with 300 mg/kgbw hydroalcoholic Anethum graveolens extract. All rats received all materials via intraperitoneal injection [IP]. 48 hours after the last IP, blood sample was taken from each animal via cardiac puncture to measure blood factors. The results indicated significant reduction in glucose, total cholesterol, triglyceride, LDL and VLDL levels in TDCG rats compared to DCG rats. Due to the use of dill extract, HDL level was significantly increased. Effect of Anethum graveolens is related to antioxidant compounds. This compounds affect glucose absorbtion and beta cells regeneration. The results of this research indicate that using hydroalcoholic extract of Anethum graveolens could effectively decrease blood sugar levels