Multicentre biobanking for rare diseases in India: Utility and challenges of lymphoblastoid cell line generation

Background and objectives For rare disease research, we aimed to establish immortalised lymphoblastoid cell lines (LCLs) from patient-derived blood samples. LCLs serve as a reliable substitute for accessing and studying the patient-specific disease biology, as patients may not always be available for repeat sampling. This study focuses on identifying the key factors that influence the successful transformation and establishment of LCLs. Methods Blood samples were collected from 187 individuals in 20 centres across 12 different cities and transported to our laboratory. Peripheral blood mononuclear cells (PBMCs) were isolated using a heparin-based density gradient method and immortalised using Epstein-Barr virus (EBV) based transformation. We sought to investigate potential correlations between success rate of transformation to LCLs and various sample-associated variables such as sample age, sample health, and ambient temperature. Results We performed the EBV transformation on 187 samples, achieving a success rate of 63%. Good quality samples achieved 80% transformation, in contrast to 45% for partially lysed samples. We noticed that samples processed within 3 d post-collection had a success rate of 71.9%, dropping to 40% for 7 d post-collection. A correlation was observed between ambient temperature and sample transformation rate, where transformation rate was highest (70%) between 31-35°C. Interpretation and conclusions Our findings identify actionable measures to improve the successful establishments of LCLs: ( i ) ensure sample quality, ( ii ) process samples within 48 h of collection, and ( iii ) maintain optimal transport conditions.