Abstract
In this contribution, we designed four types of immuno-magnetic nanoparticles for separation of different T cells (CD3+, CD4+, CD8+ and CD14+ T cells), and we established a new large-scale immuno-magnetic cell sorting procedure to achieve an enrichment of particular T cells using our designed auto-IMACS device. This device could achieve recyclable large-scale cell sorting, for which the throughput of the system reached ∼4000 mL and the maximum cell capacity was 4 × 1010. The collected cells were analyzed by flow cytometry and visual cytology data, and the effective selection rates of CD3+, CD4+, CD8+ and CD14+ T cells were 79.3%, 74.1%, 57.1% and 67.9%, respectively. The sorted CD8+ T cells still retained good cytotoxic activity against specific cells. In addition, the sorted T cells can also be further incubated in vitro and proliferated, and even could be infused back into patients for immunotherapy in the near future.
Highlights
T cells, called T lymphocytes, play a critical role in cellular immunity.[1,2,3] During immunological processes, they can be helper cells (e.g., CD4+ T cells) to assist with other white blood cells for maturation of B cells or activation of macrophages and cytotoxic T cells.[4]
The physico-chemical properties of immuno-magnetic nanoparticles (IMNPs) were investigated by measurement of magnetization loops at 300 K using the SQUID equipment (Fig. 1e)
The magnetization loops of superparamagnetic nanoparticles (SMNPs) and IMNPs based on different magnetic fields show smooth curves without hysteresis, which means that both of them present superparamagnetic behaviors
Summary
T cells, called T lymphocytes, play a critical role in cellular immunity.[1,2,3] During immunological processes, they can be helper cells (e.g., CD4+ T cells) to assist with other white blood cells for maturation of B cells or activation of macrophages and cytotoxic T cells.[4]. Further flushing steps lead to the enrichment of targeted cells in the absence of magnetic field.[13,14] This strategy has excellent specificity and separation efficiency, and some commercial products (such as CliniMACS®, Dynabeads®, MagniSortTM and Polyglobin N®) based on IMACS have been applied for different types of cells, including T cells,[15] B cells,[16] NK cells, macrophages and tumor cells.[17] while these products are commonly used for basic research with separation of small amount of cells, their high cost limits their clinical applications for large number of cells. Scheme 1 shows that this large immuno-magnetic cell sorting system could be an effective part of a entire cyclic procedure from drawing blood, targeted T cell sorting, cell proliferation to infusion back into the patient. Such a process would have vast application prospects in clinical medicine
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