The Spanish National Stem Cell Bank (Banco Nacional de Líneas Celulares, BNLC) was established in 2006. The BNLC has a network structure with 3 nodes: the Center of Regenerative Medicine in Barcelona (CMRB), the Principe Felipe Research Center (CIPF) in Valencia and the Andalusian Public Health System Biobank (SSPA Biobank) in Granada. The aim of the BNLC is to guarantee throughout the national territory the availability of human stem cell lines for biomedical research. It was established to ensure that the hESC and hiPSC generation and research are conducted according to the legal, ethical and scientific guidelines. It is mandatory to register, deposit and bank all human pluripotent cell lines (hPSC) generated in Spain and make them available to the researchers nationwide.
A number of hESC and hiPSC lines are currently registered in the BNLC (40 hESC and 146 hiPSC at 24.04.2019). Most of them have been generated by retroviral and Sendai virus infection, but there are also lines reprogrammed by lentivirus, episomal factors or mRNA. Fibroblasts were used as original cells in most of the lines, but also other cell types have been used such as cord blood, keratinocytes, peripheral blood mononuclear cells, urine cells, adipose derived mesenchymal stem cells or Schwann cells as original somatic cell lines. These cells have been donated by healthy subjects and patients suffering from more than 30 different diseases such as Parkinson’s disease, multiple sclerosis, autism, neurofibromatosis and others.
Pluripotent stem cells registered in the BNLC are fully characterized. The minimum requirements requested for characterization and QC are:
- Alkaline phosphatase test
- Pluripotency markers detection (by immunocytochemistry, flow cytometry or PCR)
Oct4, Nanog, Sox2, SSEA3, SSEA4, Tra-1-60, Tra-1-81.
- Evidence of absence of reprogramming transgenes.
- In vitro differentiation
three germ layer detection (endoderm, mesoderm and ectoderm) by embryoid bodies
formation. One marker minimum for germ layer.
- In vivo differentiation
three germ layer detection (endoderm, mesoderm and ectoderm) by teratomas induction in SCID mice. One marker minimum for germ layer.
- Mycoplasma test
- DNA fingerprinting study in the hPSC line and in the original cells.
- Confirmation of genotypic diagnosis of the cell line generated from samples with genetic mutation.
The 3 nodes of the BNLC also offer technical support for hPSC generation and characterization and even the possibility of training in handling and culture of hPSC in the facilities of the nodes. Courses and workshops about hPSC generation, culture and characterization are periodically organized.