Quality assessment on the long-term cryopreservation and nucleic acids extraction processes implemented in the andalusian public biobank.
Biobank
Cryopreservation
DNA
PBMCs
PCR
RNA
Whole blood
Journal
Cell and tissue banking
ISSN: 1573-6814
Titre abrégé: Cell Tissue Bank
Pays: Netherlands
ID NLM: 100965121
Informations de publication
Date de publication:
Jun 2019
Jun 2019
Historique:
received:
28
11
2018
accepted:
14
03
2019
pubmed:
25
3
2019
medline:
18
12
2019
entrez:
24
3
2019
Statut:
ppublish
Résumé
Human samples are commonly collected and long-term stored in biobanks for current and future analyses. Even though techniques for freezing human blood are well established, the storage time can compromise the cell viability as well as the yield and quality of nucleic acids (RNA and DNA) extracted from them. In this study, a protocol to obtain peripheral blood mononuclear cells (PBMCs) from 70 subjects, which were stored at - 196 °C from EDTA tubes for a long-term, was assessed. In parallel; a protocol to obtain DNA from the same subjects, which were stored at - 80 °C from citrate tubes, was also studied. Samples stored from 2008 to 2012 were studied and the results obtained showed that there were no statistically significant differences in the RNA or DNA extracted in terms of purity, integrity and functionality The freezing protocol used by the Málaga Biobank shows that viable PBMCs and DNA could be kept for a period of, at least, 10 years, with a high quality and performance. Furthermore, RNA extracted from these PBMCs presents also a good quality and performance. Therefore, the samples frozen according to the conditions of the protocols assessed in this study could be optimal for biomedical research.
Identifiants
pubmed: 30903409
doi: 10.1007/s10561-019-09764-9
pii: 10.1007/s10561-019-09764-9
doi:
Substances chimiques
RNA
63231-63-0
DNA
9007-49-2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
255-265Subventions
Organisme : Instituto de Salud Carlos III
ID : PT13/0010/0006
Organisme : Red Nacional de Biobancos
ID : RD09/0076/0112