Journal: Communications Medicine

Article Title: Hypothermic machine perfusion prevents hyperacute graft loss in pig-to-primate kidney xenotransplantation after 5-hours of cold Ischemia

doi: 10.1038/s43856-025-00842-6

Figure Lengend Snippet: a Representative immunohistochemical staining for CD56, CD68, and myeloperoxidase (MPO) at 2 h post-reperfusion. Upper panels show SCS-preserved kidneys; lower panels show HMP-preserved kidneys. Black line corresponds to 50 µm. b SCS-preserved kidneys in 2 h post-reperfusion had significantly higher infiltration of MPO positive neutrophils compared with HMP-preserved kidneys. There was no significant difference in CD56-positive cells (NK cells) and CD68-positive cells (macrophages) between SCS- and HMP-preserved kidneys in 1 h after transplantation. For each marker and preservation condition, five fields were analyzed per sample. Error bars indicate the standard error of the mean. HMP hypothermic machine perfusion, MPO myeloperoxidase, SCS static cold storage.

Article Snippet: To assess immune cell infiltration, immunohistochemical staining was performed using antibodies against CD56 (MA1-19129, Thermo Fisher Scientific, Waltham, MA, USA; 500:1 dilution), CD68 (sc20060, Santa Cruz Biotechnology, Dallas, TX, USA; 300:1 dilution), and myeloperoxidase (ab188211, Abcam, Cambridge, MA, USA; 1000:1 dilution).

Techniques: Immunohistochemical staining, Staining, Transplantation Assay, Marker, Preserving

Journal: Advanced Science

Article Title: Advancing Allogeneic NK Cell Immunotherapy through Microfluidic Gene Delivery

doi: 10.1002/advs.202412544

Figure Lengend Snippet: mRNA NK cell engineering and validation of cell viability and phenotype preservation. A) Confocal microscopy images of NK cells transfected with EGFP mRNA (25 µg mL −1 ), stained with 4’,6‐diamidino‐2‐phenylindole (DAPI) for nuclear visualization (scale bars: 10 µm). B) Transfection efficiency and MFI fold change for EGFP mRNA transfection in NK cells with varying mRNA concentrations ( N = three independent donors). C) Fluorescence intensity histograms of EGFP mRNA transfection in NK cells at a concentration of 25 µg mL −1 via electroporation or hydroporation ( N cell = 5000 per sample). D) Transfection efficiency, MFI fold change, and NK cell yield for EGFP mRNA transfection using electroporation or hydroporation ( N = three independent donors). E) Relative viability of hydroporated NK cells assessed with 7‐AAD staining over 6 days. F) Fluorescence intensity histograms of CD56 and CD94 immunostaining with phycoerythrin (PE)‐conjugated antibodies for the negative control and hydroporated NK cells after 24 h ( N cell = 5000 per sample). All bars represent mean ± standard deviation (SD). Student's t ‐test was used to compare the two experimental groups. n.s denotes no significant difference. Multiple comparisons were conducted using one‐way ANOVA. Student's t‐test was used to compare two experimental groups.

Article Snippet: To evaluate biomarker phenotype, PE‐conjugated monoclonal antibodies against CD56 and CD94 (Invitrogen), and NKG2A (Miltenyi Biotec) were used for immunofluorescence staining of NK cells.

Techniques: Biomarker Discovery, Preserving, Confocal Microscopy, Transfection, Staining, Fluorescence, Concentration Assay, Electroporation, Immunostaining, Negative Control, Standard Deviation