ABSTRACT
Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⺠subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.
ABSTRACT
Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⺠subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.
ABSTRACT
Subject(s)
Buffaloes , Cells, Cultured , In Vitro Techniques , Primary Cell Culture , Stem Cells , TretinoinABSTRACT
To study the effect of Cynomorium songaricum polysaccharide (CSRP) on A549 cells telomere of human non-small cell lung cancer, the mice were intragastric administrated with CSRP (0.08 g•kg⁻¹) once daily for 4 days. Then their serum was taken for preparing CSRP drug serum. A549 cells were treated by the drug serum, and the effect of drug serum with different concentrations and different treating time on the proliferation of non-small cell lung cancer A549 cells was determined by MTT test. After treating for 48 hours by the drug serum of different concentrations, the telomere length of the cells was determined by fluorescence quantitative polymerase chain reaction (qPCR); the mRNA expression of telomerase reverse transcriptase (TERT) was determined by RT-qPCR; the cells apoptosis was determined by TUNEL assay. The results demonstrated that CSRP of various concentrations could inhibit the proliferation of the lung cancer A549 cells significantly, and the inhibition effect was strongest at 48 hours with the concentration of 6.0 mL•L⁻¹. At 48 h, that CSRP of the concentrations from 1.5 to 12.0 mL•L⁻¹ could significantly shorten the telomere length of A549 cells, and the effect was strongest with the concentration of 1.5 mg•L⁻¹. CSRP of various concentrations could significantly inhibit the mRNA expression of TERT in A549 cells, and the inhibition effect was stronger when the concentration was ≥6.0 mL•L⁻¹. CSRP of various concentrations could promote A549 cells apoptosis, and the effect was stronger when the concentration was ≥6.0 mL•L⁻¹. In conclusion, CSRP has the anti-cancer effect, and the action mechanism may be associated with inhibiting TERT mRNA expression, shortening telomere length, inhibiting cells proliferation and promoting cells apoptosis.