Biobanking strategy and sample preprocessing for integrative research in monoclonal gammopathies

Varování

Publikace nespadá pod Filozofickou fakultu, ale pod Lékařskou fakultu. Oficiální stránka publikace je na webu muni.cz.

Autoři	SEVCIKOVA Tereza GROWKOVA Katerina CHYRA Zuzana FILIPOVA Jana VRUBLOVA P. JELINEK Tomas KORISTEK Z. KRYUKOV F. KRYUKOVA Elena Vladimirovna HÁJEK Roman
Rok publikování	2017
Druh	Článek v odborném periodiku
Časopis / Zdroj	Journal of clinical pathology
Fakulta / Pracoviště MU	Lékařská fakulta
Citace
www	Full Text
Doi	http://dx.doi.org/10.1136/jclinpath-2017-204329
Klíčová slova	monoclonal gammopathies; MULTIPLE-MYELOMA; BONE-MARROW; PLASMA-CELLS; CRITERIA; DISEASE; RNA
Popis	Aims Some types of monoclonal gammopathies are typified by a very limited availability of aberrant cells. Modern research use high throughput technologies and an integrated approach for detailed characterisation of abnormal cells. This strategy requires relatively high amounts of starting material which cannot be obtained from every diagnosis without causing inconvenience to the patient. The aim of this methodological paper is to reflect our long experience with laboratory work and describe the best protocols for sample collection, sorting and further preprocessing in terms of the available number of cells and intended downstream application in monoclonal gammopathies research. Potential pitfalls are also discussed. Methods Comparison and optimisation of freezing and sorting protocols for plasma cells in monoclonal gammopathies, followed by testing of various nucleic acid isolation and amplification techniques to establish a guideline for sample processing in haemato-oncology research. Results We show the average numbers of aberrant cells that can be obtained from various monoclonal gammopathies (monoclonal gammopathy of undetermined significance/light chain amyloidosis/multiple myeloma (MM)/MM circulating plasma cells/minimal residual disease MM-10 123/22 846/305 501/68 641/4000 aberrant plasma cells of 48/30/10/16/37x106 bone marrow mononuclear cells) and the expected yield of nucleic acids provided from multiple isolation kits (DNA/RNA yield from 1 to 200x10(3) cells was 2.14-427/0.12-123 ng). Conclusions Tested kits for parallel isolation deliver outputs comparable with kits specialised for just one type of molecule. We also present our positive experience with the whole genome amplification method, which can serve as a very powerful tool to gain complex information from a very small cell population.