Human renal tubular epithelial cells in serum starvation to synchronize Method.docVIP

 PAGE \* MERGEFORMAT 7 Human renal tubular epithelial cells in serum starvation to synchronize Method Author: Heli Jie, Han-Min Wang, YANG Gui-tao, Guang-Lei Chen, Wei Chen, Yang, Liu Bao, Baishu Rong, Shen Qing [Keywords:] renal tubular epithelial cells;,, synchronization;,, serum starvation;,, G0/G1 block [Abstract] Objective: To explore the use of serum starvation was established by the human renal tubular epithelial cells (HKC) synchronization method. Methods: The HKC is divided into six different low serum concentration group and control group, were used fetal calf serum concentration of 0, 1, 2, 3, 4, 5 and 100 mL / L of DMEM culture medium 24 h. According to the results of flow cytometry choose the best synchronization of FCS concentration in HKC. The synchronization of the processing time of HKC were extended to 24 h, 48 h and 72 h, to select the best synchronization time. MTT method with the observation of growth curve mapping of serum starvation on the HKC in vitro. Immunocytochemical method to verify that serum starvation synchronized with HKC effect. Results: The choice in serum-free (0 mL / L) and 1 mL / L for the best synchronization HKC concentration of fetal calf serum, choose the 48 h to synchronize the best time for the HKC. Cell growth curve shows that the use of 1 mL / L fetal bovine serum in DMEM culture medium 48 h after the return to normal cultured HKC cells, its growth curve is closer to normal cultured cells. Brdu further proves, with 1 mL / L fetal bovine serum in DMEM culture medium 48 h can make more than 90% of HKC cells in G0/G1 phase. Conclusion: 1 mL / L fetal bovine serum in DMEM culture medium 48 h access to more than 90% of the G0/G1 phase HKC. [Keywords:] renal tubular epithelial cells; synchronization; serum starvation; G0/G1 block Tubular atrophy and interstitial fibrosis is the end-stage renal disease incidence and progressive deterioration of one of the major risk factors, while the tubular atrophy and interstitial