No abstract

Although the development of gene delivery systems based on non-viral vectors is advancing, it remains a challenge to deliver plasmid DNA into human blood cells. The current “gold standard”, namely linear polyethyleneimine (l-PEI 25 kDa), in particular, is unable to produce transgene expression levels >5% in primary human B lymphocytes. Here, it is demonstrated that a well-defined 24-armed poly(2-dimethylamino) ethyl methacrylate (PDMAEMA, 755 kDa) nano-star is able to reproducibly elicit high transgene expression (40%) at sufficient residual viability (69%) in primary human B cells derived from tonsillar tissue. Moreover, our results indicate that the length of the mitogenic stimulation prior to transfection is an important parameter that must be established during the development of the transfection protocol. In our hands, four days of stimulation with rhCD40L post-thawing led to the best transfection results in terms of TE and cell survival. Most importantly, our data argue for an impact of the B cell subsets on the transfection outcomes, underlining that the complexity and heterogeneity of a given B cell population pre- and post-transfection is a critical parameter to consider in the multiparametric approach required for the implementation of the transfection protocol.

Polyelectrolyte microcapsules based on sodium cellulose sulfate (SCS) and poly‐diallyl‐dimethyl‐ammonium chloride (PDADMAC) have previously been proposed as a suitable ex vivo microenvironment for the cultivation and differentiation of primary human T lymphocytes. Here, the same system is investigated for the cultivation of human primary B cells derived from adult tonsillar tissue. Proliferation and differentiation into subtypes are followed and compared to suspension cultures of B cells from the same pool performed in parallel. Total cell expansion is somewhat lower in the capsules than in the suspension cultures. More importantly, however, the differentiation of the initially mainly memory B cells into various subtypes, in particular into plasma cell (PC), shows significant differences. Clearly, the microenvironment provided by the microcapsules is beneficial for an accelerated induction of a germinal center‐like B cell phenotype and afterward supports the long‐term survival of the PC cells. Then, varying the encapsulation conditions (i.e., presence of human serum and dedicated cytokines in the capsule core) provides a tool for finetuning the B cell response. Hence, this methodology is suggested to pave the way toward ex vivo development of human immune organoids.

Shoulder pain is regularly associated with limited mobility and limitations in activities of daily living. In occupational therapy, various interventions, including active isokinetic training with a Baltimore Therapeutic Equipment (BTE) Work Simulator, help the patient improve shoulder mobility and alleviate pain. This randomized controlled cohort study aims to evaluate the impact of different isokinetic movement patterns on the DASH score, pain, and objective performance measures, such as range of motion (ROM) and hand grip strength. Patients that participated in a specific 3-week inpatient orthopedic rehabilitation were divided into two groups. The first group (UNI-group, n = 9) carried out uniplanar exercises for shoulder flexion, abduction, and external rotation. The patients in the second group (ADL-group, n = 10) imitated multiplanar everyday movements, such as climbing on a ladder, loading a shopping cart, and raising a glass to their mouth. Compared to the UNI-group, the ADL-group improved significantly in DASH scores (mean −10.92 ± 12.59 vs. −22.83 ± 11.31), pain (NPRS −1.11 ± 2.37 vs. 3.70 ± 2.00), and shoulder abduction (+2.77 ± 15.22 vs. +25.50 ± 21.66 degrees). In conclusion, the specific BTE exercise program with multiplanar movement patterns contributed considerably to the therapeutic improvement.