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Isolation of primary human B lymphocytes from tonsils compared to blood as alternative source for ex vivo application

Titelangaben

Helm, Moritz ; Riedl, Simon ; Gollner, Katrin ; Gollner, Ulrich ; Jérôme, Valérie ; Freitag, Ruth:
Isolation of primary human B lymphocytes from tonsils compared to blood as alternative source for ex vivo application.
In: Journal of Chromatography B. Bd. 1179 (2021) . - 122853.
ISSN 1873-376X
DOI: https://doi.org/10.1016/j.jchromb.2021.122853

Angaben zu Projekten

Projektfinanzierung: Deutsche Forschungsgemeinschaft

Abstract

B lymphocytes (‘B cells’) are components of the human immune system with obvious potential for medical and
biotechnological applications. Here, we discuss the isolation of primary human B cells from both juvenile and
adult tonsillar material using a two-step procedure based on gradient centrifugation followed by separation on a
nylon wool column as alternative to the current gold standard, i.e., negative immunosorting from buffy coats by
antibody-coated magnetic beads. We show that the nylon wool separation is a low-cost method well suited to the
isolation of large amounts of primary B cells reaching purities ≥ 80%. More importantly, this method allows the
preservation of all B cell subsets, while MACS sorting seems to be biased against a certain B cell subtype, namely
the CD27 + B cells. Importantly, compared to blood, the excellent recovery yield during purification of tonsillar B
cells provides high number of cells, hence increases the number of subsequent experiments feasible with identi-
cal cell material, consequently improving comparability of results. The cultivability of the isolated B cells was
demonstrated using pokeweed mitogen (PWM) as a stimulatory substance. Our results showed for the first time
that the proliferative response of tonsillar B cells to mitogens decline with the age of the donor. Furthermore, we
observed that PWM treatment stimulates the proliferation of a dedicated subpopulation and induces some termi-
nal differentiation with ASCs signatures. Taken together this indicates that the proposed isolation procedure pre-
serves the proliferative capability as well as the differentiation capacity of the B cells.

Weitere Angaben

Publikationsform: Artikel in einer Zeitschrift
Begutachteter Beitrag: Ja
Keywords: human B lymphocytes; tonsil; blood; Nylon wool purification; MACS; Proliferation
Institutionen der Universität: Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik > Lehrstuhl Bioprozesstechnik - Univ.-Prof. Dr. Ruth Freitag
Fakultäten
Fakultäten > Fakultät für Ingenieurwissenschaften
Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik
Titel an der UBT entstanden: Ja
Themengebiete aus DDC: 500 Naturwissenschaften und Mathematik
500 Naturwissenschaften und Mathematik > 500 Naturwissenschaften
500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
600 Technik, Medizin, angewandte Wissenschaften
600 Technik, Medizin, angewandte Wissenschaften > 600 Technik
600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
Eingestellt am: 19 Jul 2021 08:14
Letzte Änderung: 10 Okt 2023 09:33
URI: https://eref.uni-bayreuth.de/id/eprint/66566