Journal: Aging (Albany NY)

Article Title: Enhanced co-culture and enrichment of human natural killer cells for the selective clearance of senescent cells

doi: 10.18632/aging.203931

Figure Lengend Snippet: Isolation and enrichment strategy of primary NK cells from human PBMC. ( A ) Experimental design of the NK cell enrichment strategy. PBMCs were collected from multiple donors (ages 20-42 years old), and NK cells were isolated and enriched. ( B ) Flow cytometry analysis of CD56 and CD16 expression in NK cells before and after enrichment for a representative donor. ( B-i ) Before enrichment, 1.64% of NK cells (0.18% of PBMCs) were CD56 Bright CD16 - and 46.34% of NK cells (2.8% of PBMCs) were CD56 Dim CD16 + NK cells. ( B-ii ) After enrichment, 2.29% of NK cells (0.47% of PBMCs) were CD56 Bright CD16 - and 66.18% of NK cells (12.6% of PBMCs) were CD56 Dim CD16 + NK cells. ( C-i ) Average percentage of CD56 Dim CD16 + NK cell population in PBMCs from five donors. ( C-ii ) Average percentage of CD56 Bright CD16 - NK cell population in PBMCs from five donors. Donor sex and age are indicated in the figure. Statistical analysis performed using paired t test. *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: Primary human arterial endothelial cells purchased from Coriell Institute for medical research (AG10770) were maintained in promo cell basal medium MV2 (PromoCell; Cat# C-22221) supplemented with Growth Medium MV 2 Supplement Pack (PromoCell; Cat# C-39221) and assayed within 10 or less passages.

Techniques: Isolation, Flow Cytometry, Expressing

Journal: Aging (Albany NY)

Article Title: Enhanced co-culture and enrichment of human natural killer cells for the selective clearance of senescent cells

doi: 10.18632/aging.203931

Figure Lengend Snippet: Activated primary NK cells selectively eliminate senescent cells. ( A ) Quantitative Realtime PCR was performed to detect the mRNA levels of CCL5 , CXCL9 , and CXCL11 in non-senescent and senescent IMR-90 fibroblasts. The results are presented as mean fold change in NS compared to S samples from two independent experiments performed in triplicate, and error bars represent ±SEM. Statistical analysis performed using unpaired t test. *p < 0.05, **p < 0.01, and ***p < 0.001. ( B ) NS or S IMR-90 fibroblasts were co-incubated with NK cells for 16 h at T:E ratios of 1:1, 1:2 and 1:3 and cytotoxicity was evaluated by LDH release. The graphs show the mean and S.E. of % LDH release. NS or S ( C-i ) doxorubicin-treated (n=6), ( C-ii ) irradiated (n=3) or ( C-iii ) etoposide-treated (n=3) IMR-90 fibroblasts or ( C-iv ) doxorubicin-treated endothelial cells (n=2) were overlayed with NK cells for 16 hours at T:E ratio of 1:1, and cytotoxicity was evaluated by LDH release. The results are plotted as mean % cytotoxicity for NS and S cells with each experiment performed in at least triplicate. The graphs show mean % LDH release. ( D ) NK cells isolated and enriched from three different individuals were co-cultured with NS or S IMR-90 cells at T:E ratio of 1:1 and cytotoxicity was evaluated by LDH release after 16 hours of co-culture. Experiments were performed in triplicate and the results are plotted as mean % cytotoxicity for NS and S. Donor sex and age are indicated in the figure. Statistical analysis performed using unpaired t test. *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: Primary human arterial endothelial cells purchased from Coriell Institute for medical research (AG10770) were maintained in promo cell basal medium MV2 (PromoCell; Cat# C-22221) supplemented with Growth Medium MV 2 Supplement Pack (PromoCell; Cat# C-39221) and assayed within 10 or less passages.

Techniques: Incubation, Irradiation, Isolation, Cell Culture, Co-Culture Assay