human nucleus pulposus cells  (Procell Inc)

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: Metabolic regulation of nucleus pulposus vells by SIN and DAB through MAPK and NF-κB pathways. (A) Volcano plot analysis of DE genes (TNF-α vs. TNF-α+SIN). (B) KEGG enrichment analysis of DE genes (TNF-α vs. TNF-α+SIN). (C) GSEA indicated MAPK signalling pathway enrichment. (D) Heatmap of ten selected differentially expressed genes (TNF-α vs. TNF-α+SIN). (E) Western blot analysis of key proteins in the MAPK signalling pathway (p-ERK, p-JNK, p-p38, ERK, JNK, and p38) in the different treatment groups. (F) Quantitative analysis of the Western blot data for MAPK signalling pathway proteins. (G–H) Western blot and quantitative analysis of p-ERK, p-JNK, p-p38, ERK, JNK, and p38 treated with or without TNF-α in the absence or presence of SIN (10 μM) (PCDNA 3.1 vs. TNFR1-OE).(I–L) Integrated analysis of TNF-α vs. TNF-α+DAB DEGs: Volcano plot of DE genes (significance/fold change), KEGG pathway enrichment of DE genes, NF-κB signalling enrichment in GSEA, and heatmap of ten selected differentially expressed genes. (M) Western blot analysis of key proteins in the NF-κB signalling pathway (p-IκBα, p-p65, IκBα, and p65) in the different treatment groups. (N) Quantitative analysis of the Western blot data for the NF-κB signalling pathway proteins. (O–P) Western blot and quantitative analysis of p-p65 and p65 (PCDNA 3.1 vs. TNFR1-OE). (Q) Matched diagram of b and y ions. (R) Western blot analysis of RELA in the different treatment groups. (S) Western blot analysis confirmed the stabilization of p65 by DAB. (T) The affinity between DAB and the p65 target protein was estimated after incubation at different temperatures via Western blot analysis. (n = 3; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001).

Article Snippet: Immortalized human nucleus pulposus cells (Procell, Cat. No. CP-H097Y) were co-transfected with an NF-κB-responsive firefly luciferase reporter plasmid and a Renilla luciferase internal control plasmid for 8 h. Following transfection, cells were treated overnight with the test compounds at a concentration of 10 μM.

Techniques: Western Blot, Incubation

Journal: Frontiers in Cell and Developmental Biology

Article Title: 17β-estradiol maintains extracellular matrix homeostasis of nucleus pulposus cells by activating p70 S6K1 signaling pathway

doi: 10.3389/fcell.2025.1564458

Figure Lengend Snippet: E2 inhibits MMPs and cleaved caspase 3 expression and reduces the ECM degradation. The rat IVD sections shown from left to right: sham ovariectomy (Sham), needle puncture plus ovariectomy with vehicle injection (OVX + Veh), and needle puncture plus ovariectomy with estradiol hormone replacement injection (OVX + E2) groups. (A, C, D) The expression of aggrecan and collagen II, which are the main components of ECM in NP tissues stained by immunohistochemical staining. (B, E, F) The expression of MMP 3 and cleaved caspase 3 in NP tissues stained by immunohistochemical staining. Values are mean ± SD (n = 5). ns: not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. IVD, intervertebral disc; OVX, ovariectomy; veh, vehicle injection; E2, 17β-estradiol; MMP, matrix metalloproteinase; ECM, extracellular matrix; NP, nucleus pulposus; SD, standard deviation.

Article Snippet: We purchased human nucleus pulposus cells (Cat NO: CP-H097) from Procell Life Science& Technology (Wuhan, China) and cultured them in DMEM/F12 (Solarbio, China) containing 10% fetal bovine serum (Gibco, Australia) and penicillin (100 U/ml)-streptomycin (100 μg/mL) (Solarbio, China) and then placed in an incubator at 37°C with 5% CO 2 .

Techniques: Expressing, Injection, Staining, Immunohistochemical staining, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: 17β-estradiol maintains extracellular matrix homeostasis of nucleus pulposus cells by activating p70 S6K1 signaling pathway

doi: 10.3389/fcell.2025.1564458

Figure Lengend Snippet: Immunohistochemistry showing that E2 mitigates IVDD by activating the p70 S6K1 signaling pathway downstream of mTOR in vivo . The rat IVD sections shown from left to right: sham ovariectomy (Sham), needle puncture plus ovariectomy with vehicle injection (OVX + Veh), and needle puncture plus ovariectomy with estradiol hormone replacement injection (OVX + E2) groups. (A) The levels of p70S6K1 and p-S6 in NP tissues of rat tails were detected using immunohistochemistry. (B, C) Data analysis of the immunohistochemistry. Values are mean ± SD (n = 5). ns: not significant; *p < 0.05; **p < 0.01; ***p < 0.001. IVD, intervertebral disc; IVDD, intervertebral disc degeneration; OVX, ovariectomy; veh, vehicle injection; E2, 17β-estradiol; NP, nucleus pulposus; p-S6, Phospho-S6 (the ribosomal protein S6); SD, standard deviation.

Article Snippet: We purchased human nucleus pulposus cells (Cat NO: CP-H097) from Procell Life Science& Technology (Wuhan, China) and cultured them in DMEM/F12 (Solarbio, China) containing 10% fetal bovine serum (Gibco, Australia) and penicillin (100 U/ml)-streptomycin (100 μg/mL) (Solarbio, China) and then placed in an incubator at 37°C with 5% CO 2 .

Techniques: Immunohistochemistry, In Vivo, Injection, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: 17β-estradiol maintains extracellular matrix homeostasis of nucleus pulposus cells by activating p70 S6K1 signaling pathway

doi: 10.3389/fcell.2025.1564458

Figure Lengend Snippet: E2 activates the p70 S6K1 signaling pathway in vitro . (A–D) Western blotting was used to analyze the levels of p70 S6K1, p-S6K1 and p-S6 in human NPCs. (E, G) The level of p70 S6K1 in human NPCs by immunofluorescence staining. (F, H) The level of p-S6 in human NPCs by immunofluorescence staining. Values are mean ± SD (n = 3). ns: not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. E2, 17β-estradiol; NPCs, nucleus pulposus cells; IL-1β, interleukin-1β; PF, PF4708671 (an p70 S6K1 inhibitor); p-S6, Phospho-S6 (the ribosomal protein S6); SD, standard deviation.

Article Snippet: We purchased human nucleus pulposus cells (Cat NO: CP-H097) from Procell Life Science& Technology (Wuhan, China) and cultured them in DMEM/F12 (Solarbio, China) containing 10% fetal bovine serum (Gibco, Australia) and penicillin (100 U/ml)-streptomycin (100 μg/mL) (Solarbio, China) and then placed in an incubator at 37°C with 5% CO 2 .

Techniques: In Vitro, Western Blot, Immunofluorescence, Staining, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: 17β-estradiol maintains extracellular matrix homeostasis of nucleus pulposus cells by activating p70 S6K1 signaling pathway

doi: 10.3389/fcell.2025.1564458

Figure Lengend Snippet: E2 activation of p70 S6K1 signaling pathway improves ECM anabolism in vitro . (A–C) Western blot analysis was used to analyze the expression of Collagen II and Aggrecan in human NPCs. (D, F) The expression of Aggrecan in human NPCs by immunofluorescence staining. (E, G) The expression of Collagen II in human NPCs by immunofluorescence staining. Values are mean ± SD (n = 3). ns: not significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. E2, 17β-estradiol; NPCs, nucleus pulposus cells; IL-1β, interleukin-1β; PF, PF4708671(an p70 S6K1 inhibitor); ECM, extracellular matrix; SD, standard deviation.

Article Snippet: We purchased human nucleus pulposus cells (Cat NO: CP-H097) from Procell Life Science& Technology (Wuhan, China) and cultured them in DMEM/F12 (Solarbio, China) containing 10% fetal bovine serum (Gibco, Australia) and penicillin (100 U/ml)-streptomycin (100 μg/mL) (Solarbio, China) and then placed in an incubator at 37°C with 5% CO 2 .

Techniques: Activation Assay, In Vitro, Western Blot, Expressing, Immunofluorescence, Staining, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: 17β-estradiol maintains extracellular matrix homeostasis of nucleus pulposus cells by activating p70 S6K1 signaling pathway

doi: 10.3389/fcell.2025.1564458

Figure Lengend Snippet: E2 activation of the p70 S6K1 signaling pathway inhibits MMP3 and cleaved caspase 3 in vitro . (A–D) Western blot analysis was used to analyze the expression of MMP3 and cleaved caspase 3. (E, F) The expression of MMP3 was determined by immunofluorescence staining. (G, H) The expression of cleaved caspase 3 was determined by immunofluorescence staining. Values are mean ± SD (n = 3). ns: not significant; *p < 0.05; **p < 0.01; ***p < 0.001. E2, 17β-estradiol; NPCs, nucleus pulposus cells; IL-1β, interleukin-1β; PF, PF4708671(an p70 S6K1 inhibitor); MMP3, matrix metalloproteinase 3; SD, standard deviation.

Article Snippet: We purchased human nucleus pulposus cells (Cat NO: CP-H097) from Procell Life Science& Technology (Wuhan, China) and cultured them in DMEM/F12 (Solarbio, China) containing 10% fetal bovine serum (Gibco, Australia) and penicillin (100 U/ml)-streptomycin (100 μg/mL) (Solarbio, China) and then placed in an incubator at 37°C with 5% CO 2 .

Techniques: Activation Assay, In Vitro, Western Blot, Expressing, Immunofluorescence, Staining, Standard Deviation

Journal: Journal of Inflammation Research

Article Title: Single-Cell Analysis Reveals Aspirin Restores Intervertebral Disc Integrity via Ferroptosis Regulation

doi: 10.2147/JIR.S519218

Figure Lengend Snippet: ( A ) Expression levels of ferroptosis inhibitory gene GPX4 and ferroptosis-activating genes CHAC1, TGFBR1, SMAD7, and CTSB at different stages. ( B ) Cell viability of human intervertebral disc nucleus pulposus (NPC) cells treated with varying concentrations (1, 5, and 10 mg/L) of LPS for 48 hours, measured by CCK-8 assay. ( C ) Representative PE fluorescence histograms of NPCs showing necrotic cells stained with PI, along with the quantitative results of necrotic percentages. ( D ) Representative fluorescent images depicting lipid peroxidation levels in NPCs, stained green with BODIPY, and nuclei stained blue with Hoechst 33342. ( E ) Representative FITC fluorescence histograms of NPCs showing cytosolic ROS production using DCFH-DA dye, with quantitative analysis of mean fluorescence intensity (MFI). *p < 0.05, ***p < 0.001 and ****p < 0.0001.

Article Snippet: Human intervertebral disc nucleus pulposus cells were purchased from Procell (#CP-H097, Wuhan, China) and passed to the third generation for experimental purposes.

Techniques: Expressing, CCK-8 Assay, Fluorescence, Staining