Table 3.
Change in the percent and MFI of CD62L on CD3−CD56+ cells from healthy donor PBMCs after enrichment and expansion. Analysis of CD62L expression on CD3−CD56+ cells before and after enrichment and expansion. PBMC populations and two-week expanded cells were stained with Pacific Blue conjugated anti-CD3 (Clone HIT3a, Biolegend), Phycoerythrin-Cyanine 7 (PE/Cy7) conjugated anti-CD56 (Clone MEM-188, Biolegend), and Allophycocyanin (APC) conjugated anti-CD62L (Clone P3.6.2.8.1, eBioscience) antibodies and fixed in 2% paraformaldehyde. For all flow cytometry experiments, appropriate IgG isotype controls were used to assess nonspecific staining. Cells were analyzed using a BD LSRII flow cytometer (BD Biosciences) and the data was analyzed using FlowJo Flow Cytometry Analysis Software (TreeStar Inc.).
| Donor | In PBMCs | In expanded NK cells on day 14 | Culture-induced change | |||||
|---|---|---|---|---|---|---|---|---|
| Percent NK cells | Percent CD62L+ NK cells | CD62L MFI of NK cells | Percent NK cell purity | Percent CD62L+ NK cells | MFI of CD62L | Change in CD62L+ NK cells after expansion | Change in CD62L+ NK cell MFI after expansion | |
| 2 | 13.1 | 30.8 | 5,647 | 94.2 | 24.8 | 4,383 | 19.5% lower | 22.4% lower |
| JL | 17.2 | 17.3 | 5,689 | 84.8 | 35.3 | 4,394 | 104.0% higher | 32.8% lower |
| 4 | 5.7 | 14.4 | 2,846 | 69.8 | 25.7 | 3,579 | 78.5% higher | 25.8% higher |
| AS | 17.0 | 68.5 | 13,316 | 95.2 | 62.9 | 9,119 | 8.2% lower | 31.5% lower |
| SS | 15.7 | 69.0 | 6,672 | 97.9 | 34.9 | 3,373 | 49.4% lower | 49.4% lower |
| *SS | 13.9 | 65.7 | 6,927 | 98.8 | 50.1 | 5,891 | 23.7% lower | 15.0% lower |
| 21 | 19.8 | 36.0 | 5,891 | 94.9 | 14.7 | 2,953 | 59.2% lower | 49.1% lower |
| *21 | 19.8 | 36.0 | 5,891 | 92.0 | 23.6 | 2,537 | 34.4% lower | 56.9% lower |
*Large-scale expansion.