| Autor: |
Remley, Victoria Ann, Jin, Jianjian, Sarkar, Sarmila, Moses, Larry, Prochazkova, Michaela, Cai, Yihua, Shao, Lipei, Liu, Hui, Fuksenko, Tatyana, Jin, Ping, Stroncek, David F., Highfill, Steven L. |
| Rok vydání: |
2021 |
| DOI: |
10.6084/m9.figshare.17079265.v1 |
| Popis: |
Additional file 1: Figure S1. T-Cell Retroviral gene transfer is enhanced using a closed-system spinoculation with CD3/CD28 Dynabeads. Schema of culture (A1) and image of centrifuge bucket orientation (B). Lymphocytes were stimulated with rIL-2 and OKT3. After 2 days in culture cell were transduced with E6 TCR retroviral vector in 6 well plates using spinoculation or bags with and without spinoculation. The transduction process was repeated on day 3. After 7 days of culture the cells were evaluated for transduction efficiency (C), fold T cell expansion (D), and cell viability (E). PBMCs enriched for lymphocyte by density gradient centrifugation were placed in PL30 bags with rIL3 and OKT3 antibody with and without CD3/CD28 Dynabeads. To simulate spinoculation, the bags were centrifuged at 1000��g for 2 h and then cultured for 7 days. Cell number (F) and cell viability (G) were measured pre-centrifugation, post-centrifugation and after 4 and 7 days of culture. Figure S2. Functional characteristics of CAR T-cells are not affected by spinoculation. Flow cytometry data from killing assay experiments at 24 (A) and 48 h (B). C) CD19/22 Healthy Donor TNF�� ELISA���s measuring cytokine levels in T-cell supernatant. D).CD19/22 Patient TNF�� ELISA���s measuring cytokine levels in T-cell supernatant. E) CD22 TNF�� ELISA���s measuring cytokine levels in T-cell supernatant. Mean and SD of triplicate wells are shown. *Indicates p ��� 0.05, ****indicates p ��� 0.0001 between the static and Sepax groups. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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