title: Biobanking strategy and sample preprocessing for integrative research in monoclonal gammopathies.
abstract: OBJECTIVE
  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 48/30/10/16/37×106bone marrow mononuclear cells) and the expected yield of nucleic acids provided from multiple isolation kits (DNA/RNA yield from 1 to 200×103cells 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.
authors.affiliation: Department of Haemato-oncology, University Hospital Ostrava, Ostrava, Czech Republic.|Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.
authors.forename: T
authors.initials: T
authors.lastname: Ševčíková
authors.affiliation: Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.|Department of Biology and Ecology, Faculty of Science, University of Ostrava, Ostrava, Czech Republic.
authors.forename: K
authors.initials: K
authors.lastname: Growková
authors.affiliation: Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.|Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Z
authors.initials: Z
authors.lastname: Kufová
authors.affiliation: Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.|Department of Biology and Ecology, Faculty of Science, University of Ostrava, Ostrava, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Filipová
authors.affiliation: Department of Haemato-oncology, University Hospital Ostrava, Ostrava, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Vrublová
authors.affiliation: Department of Haemato-oncology, University Hospital Ostrava, Ostrava, Czech Republic.|Department of Biology and Ecology, Faculty of Science, University of Ostrava, Ostrava, Czech Republic.
authors.forename: T
authors.initials: T
authors.lastname: Jelínek
authors.affiliation: Department of Haemato-oncology, University Hospital Ostrava, Ostrava, Czech Republic.|Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.
authors.forename: Z
authors.initials: Z
authors.lastname: Kořístek
authors.affiliation: Department of Haemato-oncology, University Hospital Ostrava, Ostrava, Czech Republic.
authors.forename: F
authors.initials: F
authors.lastname: Kryukov
authors.affiliation: Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.|Babak Myeloma Group, Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
authors.forename: E
authors.initials: E
authors.lastname: Kryukova
authors.affiliation: Department of Haemato-oncology, University Hospital Ostrava, Ostrava, Czech Republic.|Faculty of Medicine, University of Ostrava, Ostrava, Czech Republic.
authors.forename: R
authors.initials: R
authors.lastname: Hájek
mesh_terms: D000094902:Blood Banking / Q000379:methods; D001793:Blood Preservation / Q000379:methods*; D001800:Blood Specimen Collection / Q000379:methods; D015925:Cryopreservation / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D006801:Humans; D010265:Paraproteinemias / Q000097:blood*; D012313:RNA / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic
chemical_list: D011933:Reagent Kits, Diagnostic; D012313:RNA; D004247:DNA
keywords: DNA; HAEMATO-ONCOLOGY; METHODOLOGY; MYELOMA