The rat glomerular mesangial cell line (HBZY-1) was purchased from the China Center for Type Culture Collection. Cells were cultured in DMEM (Gibco) supplemented with 10% FBS (Gibco) and 1% penicillin-streptomycin (Gibco). To investigate the effect of MSCs on HBZY-1 stimulated by high glucose (30 mmol/L glucose for 72 h) through paracrine effects, MSCs were then plated in the bottom compartment of the Transwell chamber at a density of 400 000 cells/well, with no direct contact with HBZY-1 cells.
Hbzy 1
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
The HBZY-1 is a laboratory equipment used for cell culture applications. It is designed to provide a controlled and stable environment for the growth and maintenance of cell lines. The HBZY-1 unit regulates temperature, humidity, and atmospheric conditions to ensure optimal conditions for cell culture.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
A2494001, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by MP Biomedicals (1 mentions)
Penicillin streptomycin mixture, by BasalMedia (1 mentions)
D mannitol, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Rapamycin, by Merck Group (1 mentions)
Cox 4, by Cell Signaling Technology (1 mentions)
P mek1 2, by Cell Signaling Technology (1 mentions)
P c raf, by Cell Signaling Technology (1 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Gfp lc3 plasmid, by Addgene (1 mentions)
Z vad fmk, by Merck Group (1 mentions)
Parkin, by Cell Signaling Technology (1 mentions)
13 protocols using hbzy 1
1
Mesangial Cell-MSC Paracrine Interactions
2
Mesangial Cell Viability under Glucose and MCC950
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The rat mesangial cell line HBZY-1 was purchased from the China Center for Type Culture Collection (Wuhan, China) and were cultured in low-glucose (5.5 mmol/L) DMEM (Hyclone, SH30021, USA) with 10% FBS (Bioind, 04–001-1A, USA), 100 μg/mL streptomycin and 100 U/mL penicillin (Gibco, 15140-122, USA). The cells were routinely cultured at 5% CO2 and 37°C with saturated humidity, digested and passaged after 85% cell confluence. At 24 hrs after cell passaging and attachment, the cells were divided into five groups and cultured for 48 hrs: 1) the normal-glucose group (NG) that received 5.5 mmol/L glucose (Sigma, A24940-01,USA); 2) the high-glucose group (HG) that received 30 mmol/L glucose with vehicle (DMSO); 3) the 0.01 μM MCC950 group (0.01) that received 30 mmol/L glucose with 0.01 μmol/L MCC950; 4) the 0.1 μM MCC950 group (0.1) that received 30 mmol/L glucose with 0.1 μmol/L MCC950 and 5) the 1 μM MCC950 group (1) that received 30 mmol/L glucose with 1 μmol/L MCC950. We tested cell viability at 0 and 48 hrs by light microscopy and trypan blue exclusion.
3
Culturing Rat Glomerular Mesangial Cells
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Rat glomerular mesangial cells (HBZY-1) were purchased from the China Center for Type Culture Collection (CCTCC, Wuhan, China). HBZY-1 cells were cultured in DMEM (HyClone, Logan, UT, USA) supplemented with 10% FBS (HyClone, Logan, UT, USA) and penicillin (100 U/ml) and streptomycin (100 μg/ml). In addition, cells were maintained in a cell incubator with a humidified atmosphere of 95% air and 5% CO 2 at 37°C.
4
Culturing Rat MC Line HBZY-1 Cells
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Rat MC line HBZY-1 (China Center for Type Culture Collection, Wuhan, China) was cultured in RPMI-1640 medium. The HBZY-1 cells were cultured in a humidified 5% CO2 incubator. For the normal glucose (NG) group, cells were incubated in 5 mM glucose; for HG group, cells were incubated in 40 mM glucose.
5
Mouse MC Line HBZY-1 Cultivation and Treatment
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The mouse MC line HBZY-1 was obtained from the China Center for Type Culture Collection (CCTCC Wuhan, China). Cells were maintained at 37°C in a humidified 5% CO2 atmosphere in DMEM which contained 5.6 mM glucose, 10% fetal bovine serum (FBS; GIBCO), 100 U/mL penicillin, 100 mg/mL streptomycin, 44 mM NaHCO3, and 14 mM 4-(2-hydroxy-ethyl)-1-piperazineethanesulfonic acid. After MCs were cultivated to 60%–70% confluence, they were treated with IS for 24 h at different doses (0, 250, and 500 μM) with or without COX-2 inhibitor NS-398 treatment at a dose of 10 μM.
6
Rat Kidney Mesangial Cell Culture
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Rat kidney mesangial cells (HBZY-1) were supplied by China Center for Type Culture Collection (CCTCC, Wuhan, China), and were cultured in a humidified incubator at 37℃ containing 5% carbon dioxide using DMEM (Thermofisher, USA) supplemented containing 100 U/mL penicillin/streptomycin mixture (BasalMedia, Shanghai, China) and 10% fetal bovine serum (Thermofisher, USA). When growth to 80–90% confluence, cells were washed once with phosphate-buffered saline (PBS) and digested by 0.25% trypsin (with 0.38 g/L EDTA, BasalMedia, Shanghai, China), spare. Every fraction from R. procumbens was dissolved in DMSO (MP Biomedicals, USA) to prepare a 200 mg/mL stock solution. It was diluted to the target concentration with DMEM when used (The concentration of DMSO in each group is less than 0.5%).
7
Mesangial Cell Regulation of Collagen IV
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Rat mesangial cells (HBZY-1) were obtained from China Center for Type Culture Collection and cultured in DMEM containing 5 mM glucose and 10% fetal bovine serum. Cells were treated with 20, 25, 30, 35 and 40 mM HG for 24 h, and then 35 mM glucose plus 0.1, 0.5 and 0.9 nM C-peptide for 24 h, based on our previous study (Li et al. 2013b) . Cells were collected for detection of Col4a1-a5 mRNA expression. And after 48-h treatment, cells were collected for detection of COL4 protein content and culture medium for detection of COL4 secretion. Then, cells were divided into low glucose (LG) group (5 mM glucose), HG group (35 mM glucose) and CP group (35 mM glucose + 0.5 nM C-peptide). High l-glucose (L-HG) with the concentration of 35 mM l-glucose was used as a hypertonic control. After cells were pretreated with HG and subsequent HG plus C-peptide for 24 h, SMAD3 nuclear translocation and binding of SMAD3 to its sites in the Col4a1a2, Col4a3a4 and Col4a5 promoters were measured. The protein contents of TGFB1, TSP-1 and SMAD3 were evaluated after 48-h treatments. In addition, the localization of C-peptide was detected.
8
Rat Mesangial Cell Line Glucose Exposure
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The rat mesangial cell line HBZY-1 was purchased from the China Center for Type Culture Collection (Wuhan, China) and cultured in low-glucose Dulbecco's modified Eagle's medium (DMEM; HyClone Technologies, Logan, UT) containing 10% fetal bovine serum (HyClone Technologies). The cells were routinely cultured at 37°C and 5% CO 2 in saturated humidity. When the cells reached 90% confluence, they were assigned to three groups and eight subgroups: the normal-glucose (NG) group, in which the cells were incubated in 5.5 mmol/L D-glucose (Sigma-Aldrich, St. Louis, MO); the hypertonic control (HT) group, in which the cells were incubated in 5.5 mmol/L D-glucose DMEM + 19.5 mmol/L D-mannitol (Sigma-Aldrich); and the HG group, in which the cells were stimulated with 25 mmol/L D-glucose. All cells in the three groups were stimulated for 24 h and then cultured in 5.5 mmol/L D-glucose DMEM. At 0, 24, 48, and 72 h, the cells and their supernatants were collected.
9
Rat Mesangial Cell Culture and Cloning
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The rat mesangial cell line HBZY-1 was purchased from the China Center for Type Culture Collection (Wuhan, China), and the mesangial cells were cultured and cloned as previously described.48 (link)BSA, MTT, Z-VAD-fmk, Rapamycin, Bafilomycin A1 and U0126 were obtained from Sigma (St. Louis, MO, USA). The mRFP-GFP-LC3 plasmid was purchased from Genechem (Shanghai, China). The GFP-LC3 plasmid was kindly provided by Addgene (Cambridge, MA, USA). Primary antibodies for Caspase-3, PARP, LC3, Beclin-1, p62, p-c-Raf, total ERK1/2 (t-ERK1/2), p-ERK1/2, p-MEK1/2, t-MEK, Parkin and COX IV were purchased from Cell Signaling Technology (Danvers, MA, USA), and antibodies for Bax, Bcl-2, Caspase-9 and GAPDH were purchased from Abcam (Cambridge, MA, USA).
10
Rat Mesangial Cell Line HBZY-1 Culture
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The rat mesangial cell line HBZY-1 was purchased from the China Center for Type Culture Collection and cultured in low-glucose Dulbecco’s modified Eagle’s medium (DMEM; Hyclone,USA) containing 10% fetal bovine serum (FBS; Hyclone,USA), 100 U/ml penicillin, and 100 U/ml streptomycin. The cells were routinely cultured at 37°C and 5% CO2 with saturated humidity. The cells were digested and passaged when the cell confluence was greater than 85%. At 24 h after passaging and cell attachment, the cells were divided into the following groups: A, the normal-glucose group (NG), which received 5.5 mmol/L glucose (Sigma); B, the hypertonic control group (MA), which received 5.5 mmol/L glucose+19.5 mmol/L mannitol (Sigma); C, the high-glucose group (HG), which received 25 mmol/L glucose; D, the LY294002 intervention group (LY294002), which received 25 mmol/L glucose+5 μmol/L LY294002 (CST; Sigma); and E, the UA intervention group (UA), which received 25 mmol/L glucose+2.5 μmol/L UA (Sigma).
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