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

Title data

Helm, Moritz ; Riedl, Simon ; Gollner, K. ; Gollner, U. ; 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. Vol. 1179 (2021) . - No. 122853.
ISSN 1873-376X
DOI: https://doi.org/10.1016/j.jchromb.2021.122853

Project information

Project financing: Deutsche Forschungsgemeinschaft

Abstract in another language

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.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: human B lymphocytes; tonsil; blood; Nylon wool purification; MACS; Proliferation
Institutions of the University: Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Faculties
Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Result of work at the UBT: Yes
DDC Subjects: 500 Science
500 Science > 500 Natural sciences
500 Science > 570 Life sciences, biology
600 Technology, medicine, applied sciences
600 Technology, medicine, applied sciences > 600 Technology
600 Technology, medicine, applied sciences > 610 Medicine and health
Date Deposited: 19 Jul 2021 08:14
Last Modified: 20 Jul 2021 06:07
URI: https://eref.uni-bayreuth.de/id/eprint/66566