Optimizing in vitro T cell differentiation by using induced pluripotent stem cells with GFP-RUNX1 and mCherry-TCF7 labelling.

In vitro T-cell differentiation from pluripotent stem cells (PSCs) could potentially provide an unlimited source of T cells for cancer immunotherapy, which, however is still hindered by the inefficient obtaining functionally-matured, terminally-differentiated T cells. Here, we established a fluorescence reporter human induced pluripotent stem cell (iPSC) line termed TCF7mCherryRUNX1GFP, in which the endogenous expression of RUNX1 and TCF7 are illustrated by the GFP and mCherry fluorescence, respectively. Utilizing TCF7mCherryRUNX1GFP, we defined that the feeder cells incorporating CXCL12-expressing OP9 cells with DL4-expressing OP9 cells at a 1:3 ratio (OP9-C1D3) significantly enhanced efficiency of CD8+ T cell differentiation from PSCs. Additionally, we engineered a chimeric antigen receptor (CAR) targeting EGFR into iPSCs. The CAR-T cells differentiated from these iPSCs using OP9-C1D3 feeders demonstrated effective cytotoxicity toward lung cancer cells. We anticipate this platform will help the in vitro HSPC and T cell differentiation optimization, serving the clinical demands of these cells.

Variation in Glucosinolate Accumulation among Different Sprout and Seedling Stages of Broccoli (Brassica oleracea var. italica).

Glucosinolates (GLs) are plant secondary metabolites that may act against different types of cancers. Broccoli (Brassica oleracea var. italica) is rich in GLs which makes it an excellent source of these nutraceuticals. The composition and concentration of GLs vary among broccoli cultivars and throughout the developmental stages of the plant. To obtain the GL profiles of broccoli, GL compositions and contents in four early developmental stages (seeds, 3-day sprouts, 11-day and 17-day seedlings) were determined for nine cultivars of broccoli in this study. A total of 12 GLs including 9 aliphatic GLs and 3 indole GLs were identified from the nine broccoli cultivars using LC-QTOF-MS. UPLC results showed that aliphatic GLs concentrations decreased with broccoli sprouts and seedling growth for most cultivars. Interestingly, indole GLs amounts increased after germination and reached the highest level in 3-day sprouts or 11-day seedlings, and they fell back to a low level in 17-day seedlings. The GL profiles of nine cultivars documented in this study will provide useful information for high quality germplasm selection for cultivation or genetic engineering, and further understanding of the GL metabolic pathways.