Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel cultivation strategy to recover NK cell cytotoxicity

doi: 10.3389/fbioe.2026.1797129

Figure Lengend Snippet: Total VCC, viability (A) , cytotoxicity, specific growth rate (B) , cytotoxic capacity (C) , lactate concentration and production rate (D) , glucose concentration and consumption rate (E) , glutamine concentration and consumption rate (F) of the NK-92 cell batch expansion process in static T-flasks. Error bars represent the standard deviation of three, independent biological replicates (n = 3).

Article Snippet: To assess the cytotoxicity of the NK-92 cells, K-562-GFP (K562) cells (CCL-243-GFP, American Type Culture Collection, Manassas, VA, United States) were maintained at standard cultivation conditions in a Roswell Park Memorial Institute 1640 medium (RPMI; Gibco, Grand Island, NY, United States) containing 5% heat-inactivated fetal bovine serum (FBS, Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Concentration Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel cultivation strategy to recover NK cell cytotoxicity

doi: 10.3389/fbioe.2026.1797129

Figure Lengend Snippet: (A) Graphical overview of the design space used to determine the cytotoxicity recovery without an additional factor of recovery duration. Overview of significant factors used for the calculation of the PLS model for cytotoxicity (B) and cytotoxic capacity (C) after normalization of the coefficients. Only significant factors are displayed (different than zero) with a standard deviation smaller than the factor’s value - Med: medium composition, Temp: cultivation temperature Dur: recovery duration. (D,E) Contour plots illustrating PLS-based predictions of the optimal recovery setpoint, derived from the analysis of DoE responses for cytotoxicity and cytotoxic cells. The circles on both plots represent the combined optimum conditions that maximize cytotoxicity and cytotoxic capacity recovery. The factor recovery duration was set to the identified optimum of 3.7 days.

Article Snippet: To assess the cytotoxicity of the NK-92 cells, K-562-GFP (K562) cells (CCL-243-GFP, American Type Culture Collection, Manassas, VA, United States) were maintained at standard cultivation conditions in a Roswell Park Memorial Institute 1640 medium (RPMI; Gibco, Grand Island, NY, United States) containing 5% heat-inactivated fetal bovine serum (FBS, Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Standard Deviation, Derivative Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel cultivation strategy to recover NK cell cytotoxicity

doi: 10.3389/fbioe.2026.1797129

Figure Lengend Snippet: Time-resolved measurements of cytotoxicity (A) and cytotoxic capacity (B) during the cytotoxicity recovery process. Cytotoxicity (C) , cytotoxic capacity (D) , total viable cell count (E) , viability (F) , cumulative population doublings (G) , and STY (H) were compared between medium compositions on day 3 when cytotoxicity recovery reached its maximum, using one-way ANOVA, where p ≥ 0.05 is not significant (ns), p < 0.05 is *, p < 0.01 is **, p < 0.001 is ***, and p < 0.0001 is ****. Error bars represent standard deviations of all runs per medium composition (n = 4 for fresh medium (green), n = 8 for mixed medium (red) and n = 4 for spent medium (blue)). Correlation between cytotoxicity at day 3 of the recovery and lactate concentration (I) and pH (J) at day 0 of cytotoxicity recovery. Regression was calculated with simple linear regression and correlation coefficients were calculated with Pearson’s correlation.

Article Snippet: To assess the cytotoxicity of the NK-92 cells, K-562-GFP (K562) cells (CCL-243-GFP, American Type Culture Collection, Manassas, VA, United States) were maintained at standard cultivation conditions in a Roswell Park Memorial Institute 1640 medium (RPMI; Gibco, Grand Island, NY, United States) containing 5% heat-inactivated fetal bovine serum (FBS, Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Cell Characterization, Concentration Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: A novel cultivation strategy to recover NK cell cytotoxicity

doi: 10.3389/fbioe.2026.1797129

Figure Lengend Snippet: Total viable cell count and viability (A) , cytotoxicity and lactate concentration (C) , and cytotoxic capacity and specific growth rate (E) of NK-92 cells during a fed-batch expansion process in a 2-L working volume stirred-tank bioreactor. Total VCC and viability (B) , cytotoxicity and lactate concentration (D) , and cytotoxic capacity and specific growth rate (F) of NK-92 cells during the cytotoxicity recovery phase following expansion in the same bioreactor are shown. Due to the limited working volume of the bioreactor, only 6.6% of all cells produced during the fed-batch phase could be retained for the recovery phase (hence, different absolute values of the total viable cell count and cytotoxic capacity are displayed). Viablity at harvest (G) and STY at harvest (H) after the expansion process phase and the recovery process phase were compared uising an unpaired t-test where p ≥ 0.05 is not significant (ns), p < 0.05 is *, p < 0.01 is **, p < 0.001 is ***, and p < 0.0001 is ****. Error bars for expansion process and recovery process represent technical triplicates (n = 3).

Article Snippet: To assess the cytotoxicity of the NK-92 cells, K-562-GFP (K562) cells (CCL-243-GFP, American Type Culture Collection, Manassas, VA, United States) were maintained at standard cultivation conditions in a Roswell Park Memorial Institute 1640 medium (RPMI; Gibco, Grand Island, NY, United States) containing 5% heat-inactivated fetal bovine serum (FBS, Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Cell Characterization, Concentration Assay, Produced

Journal: Pharmaceutics

Article Title: Modulation of the Cytotoxic Properties of Pd(II) Complexes Based on Functionalized Carboxamides Featuring Labile Phosphoryl Coordination Sites

doi: 10.3390/pharmaceutics15041088

Figure Lengend Snippet: Cytotoxicity of the phosphoryl-functionalized amide derivatives against some human hematopoietic c ancer cell lines.

Article Snippet: To study the apoptosis inducing ability of complex 3b , K562 and K562/iS9 cells, preincubated for a day in a CO 2 incubator at 37 °C, were cultured in the medium containing 10 μM of the palladocycle for 20 h. After exposure, the cells were washed with cold PBS and incubated with Annexin V-FITC for 20 min before being treated with PI according to the supplier protocol (Elabscience Annexin V-FITC/PI Apoptosis Detection Kit).

Techniques:

Journal: Pharmaceutics

Article Title: Modulation of the Cytotoxic Properties of Pd(II) Complexes Based on Functionalized Carboxamides Featuring Labile Phosphoryl Coordination Sites

doi: 10.3390/pharmaceutics15041088

Figure Lengend Snippet: Percentages of necrotic (upper left), early apoptotic (lower right), and late apoptotic (upper right) K562 ( a , b ) and K562/iS9 ( c , d ) cells in the control experiments ( a , c ) and after exposure to complex 3b ( b , d ) for 20 h.

Article Snippet: To study the apoptosis inducing ability of complex 3b , K562 and K562/iS9 cells, preincubated for a day in a CO 2 incubator at 37 °C, were cultured in the medium containing 10 μM of the palladocycle for 20 h. After exposure, the cells were washed with cold PBS and incubated with Annexin V-FITC for 20 min before being treated with PI according to the supplier protocol (Elabscience Annexin V-FITC/PI Apoptosis Detection Kit).

Techniques: Control

Journal: RMD Open

Article Title: Development of an enzyme-linked immunosorbent assay for the efficient detection of autoantibodies against nuclear valosin-containing protein-like protein (NVL) 2 using its manipulated cDNA

doi: 10.1136/rmdopen-2025-005679

Figure Lengend Snippet: Representative results of IP-WB using K562 nuclear extract for anti-NVL2 antibodies and an indirect immunofluorescence image for an anti-NVL2 antibody-positive serum sample. ( A ) Immunoprecipitates from K562 nuclear extracts with patient’s serum were probed with anti-NVL polyclonal antibody to confirm the ELISA results using the recombinant protein. Of the 28 ELISA high-titre sera, 18 sera showed reactivity with an approximately 95 kDa band by IP-WB. Lane 1 shows an input lane that contains half the dose of K562 extract used. Lanes 2, 3 and 4 correspond to Pt. 1, 7 and 11 in , respectively. Lanes 5 and 6 correspond to IP-WB negative patients among the ELISA high-titre sera. Lane 7 is serum from a healthy control. An approximately 95 kDa band was detected in lanes 2–4, but not in lanes 5–7. Lane 7 is serum from a healthy control. *denotes the position of 95 kDa molecular weight proteins in the IP-WB analysis. ( B ) In the AC-8 pattern seen by indirect immunofluorescence on anti-NVL2 antibody-positive serum sample, we see diffuse fluorescence of the entire nucleolus (blue arrowhead) and no staining of the metaphase plate (red arrowhead). IP, immunoprecipitation; NVL, nuclear valosin-containing protein-like protein; Pt, patient; WB, Western blotting

Article Snippet: IP-WB was performed using K562 nuclear cell extracts (Santa Cruz Biotechnology, Dallas, Texas, USA) without cross-linking, as described previously, with minor modifications.

Techniques: Immunofluorescence, Enzyme-linked Immunosorbent Assay, Recombinant, Control, Molecular Weight, Fluorescence, Staining, Immunoprecipitation, Western Blot

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: The inhibitory activity of Jac-A on tumour cells via MTT Assay (IC 50 , μ M, n = 4, mean ± SD)

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Activity Assay, MTT Assay

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A caused K562 cell apoptosis. 10 6 of K562 cells were treated with different concentrations of Jac-A for 48 h. Cells were then stained with Annexin V-FITC and propidium iodide, and analysed using flow cytometry. Four different cell populations marked as the following: live cell population (PI - AV-), early apoptosis (PI - AV+), late apoptosis (PI + AV+) and dead cells (PI + AV-). Cells were treated with no Jac-A (A) , 0.5% DMSO (B) , or 0.1, 1, 5, 10 μM/L Jac-A (C, D, E and F) . The results are one representative of three independent experiments.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Staining, Flow Cytometry

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A induced caspase-dependent apoptosis in K562 cells. (A) Cyt c release in Jac-A-induced apoptosis of K562 cells. 10 6 of K562 cells were treated with different concentration of Jac-A for 48 h. Cytosol and mitochondrial heavy membrane samples were prepared and subjected to immunoblot with anti-Cyt c specific antibodie as described in Materials and methods. The results are one representative of three independent experiments. (B) Western blot showing conspicuous cleavage of caspase-3, caspase-9, and PARP in K562 cells treated with Jac-A. K562 cells subjected to protein extract preparation after treated with 0 (control), 3, 6, 12 μM/L Jac-A for 48 h. Then, fifty micrograms extracted protein subjected to Western blot using anti-PARP, PARP, cleaved caspase-9, caspase-9, cleaved caspase-3, caspase-3, and β-actin. The results are one representative of three independent experiments. (C) Jac-A-induced apoptosis was inhibited in a concentration dependent manner by the Z-VAD-fmk. K562 cells were first treated with or without different concentrations of Z-VAD-fmk for 4 h, followed by the treatment of Jac-A (10 μM) for 48 h.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Concentration Assay, Membrane, Western Blot, Control

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A inhibited the heterodimerization of antiapoptotic proteins with pro-apoptotic proteins. Jac-A inhibited the binding between Bax with Bcl-x L or Bcl-2 (A) and Bak with Mcl-1 (B) . K562 cells were treated with indicated concentrations of Jac-A for 48 h. 150 μg of K562 cell lysates were subjected to co-immunoprecipitation using anti-Bax and anti-Bak antibodies, respectively, further immunoblot with anti- Bcl-x L , Bcl-2, Mcl-1, Bax, or Bak antibody, as described in Materials and methods. The results are one representative of three independent experiments.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Binding Assay, Immunoprecipitation, Western Blot

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Therapeutic study of Jac-A in the K562-bearing mice (n = 10/group). (A) Tumour volume plot of K562-bearing mice treated with vehicle or Jac-A at 2, 10, or 50 mg/kg by oral gavage for 21 days. The tumours were measured twice per week. The data are represented as the mean ± SEM. Tumour growth was inhibited significantly after treatment with Jac-A compared with the control group. *, P < 0.05; †, P < 0.01; ‡, P < 0.001 compared with the control group. (B) Selected nude mice models of different groups treated with Jac-A or vehicle at day 14 after therapy. (C) Sizes of selected tumours harvested from dead nude mice bearing K562 cells from different groups treated with the vehicle or Jac-A. (D) Kaplan-Meier survival plot of the K562-bearing nude mice. The survival of the K562-bearing nude mice was prolonged in the Jac-A treated groups compared with control group. (E) Body weight plot of the K562-bearing nude mice. The data are represented as the mean ± SEM. *, P < 0.05; †, P < 0.01; ‡, P < 0.001 compared with the control group.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Control