Production of GMP-grade human UC-MSCs
human T lymphocytic virus
STERILE, VIRUS, HEPATITIS C, MYCOPLASMA
TH-SC01 is a cellular product manufactured in a Good Manufacturing Practice (GMP) facility at TopCel-KH Biopharmaceutical (Nanjing, China). Umbilical cord tissue was donated by a healthy volunteer whose baby was delivered by elective cesarean section. Informed consent was obtained before cesarean section.Briefly, cord tissue was placed in a sterile bottle containing DMEM/F12 and transported to the GMP facility. After umbilical vessels were removed, Wharton’s jelly was sliced into fragments of about 1 cm in length. The fragments were placed on culture plates in DMEM/F12 media with 1% platelet lysate (AventaCell BioMedical Corp, Atlanta, USA). After about 15 days, cord fragments were discarded and cells were digested with GMP-grade enzymes (Gibco Life Technologies, Waltham, USA). Cells were subsequently passaged. UC-MSCs of passage 3 were cryopreserved in cryovials in the vapor phase of liquid nitrogen as master cell bank (MCB). For quality control, karyotyping and viral tests were performed. Karyotype analysis confirmed a normal human karyotype. UC-MSCs were also evaluated for cell surface markers CD44, CD73, CD90, CD105, CD31, CD34, and CD45. We thawed cells for subculture under the same conditions. Cells cultured to passage 5 were used in the present study.The final cells were harvested into a solution with 5% human serum albumin. Release testing included total nucleated cell count, viability, endotoxin, and pathogens (human immunodeficiency virus-1 and -2, cytomegalovirus, hepatitis B virus, hepatitis C virus, human T lymphocytic virus, Epstein-Barr virus, and mycoplasma). Pathogens were detected using reverse transcriptase PCR. For each patient, 120 million cells were formulated in 24 mL and supplied in four vials (6 mL/vial). Cells were shipped to the hospital on the day of administration. The formulated product can be stored for a maximum of 48 h between 4 and 8 °C.