Immunomagnetic isolation of myeloma cells by autoMACS

k 2007

Immunomagnetic isolation of myeloma cells by autoMACS

ZELENÁ, Jana; Darina OČADLÍKOVÁ; Drahomíra KYJOVSKÁ; Jana MORAVCOVÁ; Marcela RYCOVÁ et. al.

Základní údaje

Originální název

Immunomagnetic isolation of myeloma cells by autoMACS

Název česky

Imunomagnetická isolace myelomových buněk pomocí přístroje autoMACS

Autoři

Vydání

From cell sorting to plasma cell identification and chromosomal aberrations in multiple myeloma and Immunomagnetic cell separation for clinical use – Application manual, 2007

Další údaje

Jazyk

angličtina

Typ výsledku

Prezentace na konferencích

Obor

Genetika a molekulární biologie

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Lékařská fakulta

ISBN

978-80-210-4417

Klíčová slova anglicky

immunomagnetic cell separation; plasma cells; clinical use

Štítky

clinical use, immunomagnetic cell separation, plasma cells

Příznaky

Mezinárodní význam

Změněno: 21. 5. 2008 11:39, Mgr. Jana Čumová, Ph.D.

Anotace

ORIG CZ

V originále

The standard method used for cell separation is Magnetic Antibody Cell Separation, MACS. In our laboratories, we use autoMACS separator (Miltenyi Biotec, Bergisch Gladbach, Germany) designed for positive selection as well as depletion of magnetically labeled cells. The separator operates with separation programs according to cell abundance and the intensity of marker expression. The separated cells are immediately ready for experiments, cell analysis or further subset sorting. Within minutes can be isolated up to 2x108 pure target cells per sample. AutoMACS allows positive selection and depletion of cells even if the antigen used for selection is expressed at low levels. CD138+ myeloma cells can be isolated from whole blood or bone marrow (BM) or from mononuclear cells prepared using gradient centrifugation from BM of the patients with multiple myeloma. The CD138 antigen, also known as syndecan-1, is found on malignant plasma cells in the BM. The CD138+ myeloma cells are isolated via positive selection of human plasma cells using CD138 micro beads. The sample containing labeled cells is loaded onto an autoMACS column which is placed in a magnetic field. The magnetically labeled CD138+ cells are retained within the column. The unlabeled cells run through as negative fraction. Isolated plasma cells can be used for molecular biology studies such as PCR analysis, microarrays, and protein analysis, and for analysis of cell surface markers of intracellular antigens, for example, immunoglobulin light chains. Using possel d program, we obtained 77,6% cells (8,9-112,4%), average (min-max), with purity of positive fraction 83,8% (63,7-98,9%). In this report, we would like to inform about our achieved results in immunomagnetic separation of myeloma cells from patients using several separation programs.

Česky

Přístroj autoMACS (Miltenyi Biotec, Bergisch Gladbach, Německo) umožňuje separaci plazmatických buněk (CD138+) z periferní krve a kostní dřeně pacientů nemocných mnohočetným myelomem. Značené myelomové buňky protilátkou anti-human CD138+ konjugovanou s paramagnetickou kuličkou jsou separovány z KD pomocí pozitivní strategie na novém přístroji autoMACS. V naší práci prezentujeme první výsledky separací plazmatických buněk pomocí různých separačních programů přístroje autoMACS (possel d, possel d2, possel s).

Návaznosti

LC06027, projekt VaV

Název: Univerzitní výzkumné centrum - Česká myelomová skupina (Akronym: LC MGUS)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Univerzitní výzkumné centrum - Česká myelomová skupina (URC - CMG)

Citovat

ZELENÁ, Jana; Darina OČADLÍKOVÁ; Drahomíra KYJOVSKÁ; Jana MORAVCOVÁ; Marcela RYCOVÁ; Lucie KOVÁŘOVÁ; Luděk POUR; Renata SUSKÁ; Petra VIDLÁKOVÁ; Božena HANÁKOVÁ; Marta KREJČÍ; Zdeněk ADAM a Roman HÁJEK. Immunomagnetic isolation of myeloma cells by autoMACS. In From cell sorting to plasma cell identification and chromosomal aberrations in multiple myeloma and Immunomagnetic cell separation for clinical use – Application manual. 2007. ISBN 978-80-210-4417.

@proceedings{727471,
   author = {Zelená, Jana and Očadlíková, Darina and Kyjovská, Drahomíra and Moravcová, Jana and Rycová, Marcela and Kovářová, Lucie and Pour, Luděk and Suská, Renata and Vidláková, Petra and Hanáková, Božena and Krejčí, Marta and Adam, Zdeněk and Hájek, Roman},
   booktitle = {From cell sorting to plasma cell identification and chromosomal aberrations in multiple myeloma and Immunomagnetic cell separation for clinical use – Application manual},
   keywords = {immunomagnetic cell separation; plasma cells; clinical use},
   language = {eng},
   isbn = {978-80-210-4417},
   title = {Immunomagnetic isolation of myeloma cells by autoMACS},
   year = {2007}
}

TY  - CONF
ID  - 727471
AU  - Zelená, Jana - Očadlíková, Darina - Kyjovská, Drahomíra - Moravcová, Jana - Rycová, Marcela - Kovářová, Lucie - Pour, Luděk - Suská, Renata - Vidláková, Petra - Hanáková, Božena - Krejčí, Marta - Adam, Zdeněk - Hájek, Roman
PY  - 2007
TI  - Immunomagnetic isolation of myeloma cells by autoMACS
SN  - 978802104417
KW  - immunomagnetic cell separation
KW  - plasma cells
KW  - clinical use
N2  - The standard method used for cell separation is Magnetic Antibody Cell Separation, MACS. In our laboratories, we use autoMACS separator (Miltenyi Biotec, Bergisch Gladbach, Germany) designed for positive selection as well as depletion of magnetically labeled cells. The separator operates with separation programs according to cell abundance and the intensity of marker expression. The separated cells are immediately ready for experiments, cell analysis or further subset sorting. Within minutes can be isolated up to 2x108 pure target cells per sample. AutoMACS allows positive selection and depletion of cells even if the antigen used for selection is expressed at low levels. CD138+ myeloma cells can be isolated from whole blood or bone marrow (BM) or from mononuclear cells prepared using gradient centrifugation from BM of the patients with multiple myeloma. The CD138 antigen, also known as syndecan-1, is found on malignant plasma cells in the BM. The CD138+ myeloma cells are isolated via positive selection of human plasma cells using CD138 micro beads. The sample containing labeled cells is loaded onto an autoMACS column which is placed in a magnetic field. The magnetically labeled CD138+ cells are retained within the column. The unlabeled cells run through as negative fraction. Isolated plasma cells can be used for molecular biology studies such as PCR analysis, microarrays, and protein analysis, and for analysis of cell surface markers of intracellular antigens, for example, immunoglobulin light chains. Using possel d program, we obtained 77,6% cells (8,9-112,4%), average (min-max), with purity of positive fraction 83,8% (63,7-98,9%). In this report, we would like to inform about our achieved results in immunomagnetic separation of myeloma cells from patients using several separation programs.
ER  -

ZELENÁ, Jana; Darina OČADLÍKOVÁ; Drahomíra KYJOVSKÁ; Jana MORAVCOVÁ; Marcela RYCOVÁ; Lucie KOVÁŘOVÁ; Luděk POUR; Renata SUSKÁ; Petra VIDLÁKOVÁ; Božena HANÁKOVÁ; Marta KREJČÍ; Zdeněk ADAM a Roman HÁJEK. Immunomagnetic isolation of myeloma cells by autoMACS. In \textit{From cell sorting to plasma cell identification and chromosomal aberrations in multiple myeloma and Immunomagnetic cell separation for clinical use – Application manual}. 2007. ISBN~978-80-210-4417.

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