Trypsin edta solution a

CAFs were isolated from the initial passages of PDTO cultures. CAFs were identified microscopically by their spindle-like appearance, and whenever outgrowth was observed, the dome was left in the well when PDTOs were passaged to allow CAFs to expand and attach to the bottom of the plate. After CAF expansion to approximately 80% confluence of the well, the medium was aspirated, and 300 µL Trypsin/EDTA Solution A (0.25%/0.02%) (03-050-1A, Biological Industries, Beit Haemek, Israel) was added to the well for 1–3 min under close observation under the microscope. CAFs would detach from the plastic surface sooner than the tumor cells, allowing them to be separated and collected in the aspirate.
Once CAFs had been isolated, they were cultured in T75 plastic flasks in 89% RPMI 1640 (61870010, Gibco, ThermoFisher), 10% Fetal Bovine Serum (FBS) (A4766801, ThermoFisher), and 1% penicillin/streptomycin 10,000 U/mL (15140122, ThermoFisher) and passaged once 80% confluent.

IR780 was obtained from Sigma-Aldrich and used as received. TPZ was obtained from MedChemExpress and used as received. All other chemicals were purchased from Chongqing Chuandong Chemical (group) CO., Ltd. and used without further purification. All chemicals were analytical grade and used without further purification if not indicated otherwise. Dulbecco’s modified eagle’s medium (DMEM) was purchased from GE Healthcare Life Sciences HyClone Laboratories (HyClone, USA). Fetal bovine serum (FBS) was purchased from VivaCell (Shanghai, China), and Trypsin EDTA Solution A was purchased from Biological Industries (BI, Israel). Cell Counting kit-8 (CCK-8) was purchased from Fdbio science CO., Ltd. (Hangzhou, China) and used as received. Dialysis tubes were purchased from Shanghai Lvniao Technology Group. Clear polystyrene tissue culture treated 6-well and 96-well plates were purchased from JET BIOFIL CO., Ltd. (Guangzhou, China). The deionized water was used in the experiment, which was purified from Milli-Q (Millipore, 18.2 MΩ cm⁻¹).

HEK293, HEK293T, ACE2-expressing
HEK293T cells, and stable pAGDisplay-RBD cell lines were cultured
at 37 °C in a 5% CO₂ atmosphere in DMEM medium (4.5
g/L glucose, l-glutamine; Gibco, USA) supplemented with 10%
fetal bovine serum (FBS) and 1% glutamine (4 mM). Cells were passaged
2–3 times per week using trypsin EDTA solution A (Biological
Industries, USA) for cell detachment. The stably expressing ACE2 HEK293T
cell line was kindly obtained from the lab of Dr. Ron Diskin (Weizmann
Institute of Science) and kept under puromycin antibiotics (0.5 μg/mL,
Invitrogen, USA).