Human-induced pluripotent stem cell-derived macrophages and their immunological function in response to tuberculosis infection

Abstract

BackgroundInduced pluripotent stem cells (iPS) represent an innovative source for the standardized in vitro generation of macrophages (Mφ). Mφ show great promise in disease pathogenesis, particularly tuberculosis. However, there is no information about human iPS-derived (hiPS) macrophages (hiPS-Mφ) in response to tuberculosis infection.MethodsIn the present study, macrophages derived from hiPS were established via embryoid body (EB) formation by using feeder-free culture conditions, and the human monocyte cell line THP-1 (THP-1-Mφ) was used as control. iPS-Mφ were characterized by using morphology, Giemsa staining, nonspecific esterase staining (α-NAE), phagocytosis, and surface phenotype. Additionally, after treatment with Bacillus Calmette-Guérin (BCG) for 24 h, cell apoptosis was detected by using an Annexin V-FITC Apoptosis Detection assay. The production of nitric oxide (NO), expression of tumor necrosis factor alpha (TNF-α), activity of apoptosis-related protein cysteine-3 (Caspase-3) and expression of B-cell lymphoma-2 (Bcl-2) were analyzed.ResultsWith respect to morphology, surface phenotype, and function, the iPS-Mφ closely resembled their counterparts generated in vitro from a human monocyte cell line. iPS-Mφ exhibited the typically morphological characteristics of macrophages, such as round, oval, fusiform and irregular characteristics. The cells were Giemsa-stained-positive, α-NAE-positive, and possessed phagocytic ability. iPS-Mφ express high levels of CD14, CD11b, CD40, CD68, and major histocompatibility complex II (MHC-II). Moreover, with regard to the apoptotic rate, the production of NO, expression of TNF-α, and activity of Caspase-3 and Bcl-2, iPS-Mφ closely resemble that of their counterparts generated in vitro from human monocyte cell line in response to BCG infection. The rate of apoptosis of BCG-treated iPS-Mφ was 37.77 ± 7.94% compared to that of the untreated group at 4.97 ± 1.60% (P < 0.01) by using Annexin V-FITC Apoptosis Detection. Additionally, the rate of apoptosis of BCG-treated THP-1-Mφ was 37.1 ± 2.84% compared to that of the untreated group at 6.19 ± 1.68% (P < 0.001). The expression of TNF-α and the production of NO were significantly increased (P < 0.001), and the activity of Caspase-3 was increased. However, the expression of Bcl-2 was inhibited (P < 0.001).ConclusionsOur results demonstrate that Mφ derived from hiPS perform the immunological function in response to Bacillus Calmette-Guérin infection by undergoing apoptosis, increasing the production of NO and expression of TNF-α. Thus, our study may help to overcome the limitations of research into certain rare diseases due to the lack of adequate supply of disease-specific primary cells.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0800-x) contains supplementary material, which is available to authorized users.

THP-1 and Dictyostelium Infection Models for Screening and Characterization of Anti-Mycobacterium abscessus Hit Compounds

“…For infection studies, we used the THP-1 monocyte cell line, cells of which were differentiated with phorbol 12-myristate 13-acetate (PMA) and infected with M. abscessus at a multiplicity of infection (MOI) of 5:1 in batch. THP-1 cells were used mainly for their uniformity, their similar behavior to macrophages derived from human stem cells (20), and the option to compare our studies to those described in the literature (19,21). The infected cell suspension was dispensed into 96-well plates.…”

Macrophages as a Double-Edged Weapon: The Use of Macrophages in Cancer Immunotherapy and Understanding the Cross-Talk Between Macrophages and Cancer

“…In our laboratory, we optimized a protocol to derive Mfs from iPSCs through the embryoid bodies (EBs) method in which iPSCs aggregate to form three-dimensional-like structures called EBs that have the three germ layers. Then, the EBs are induced toward the hematopoietic lineage to produce Mfs (Hong et al, 2018). The advantage of using stem cells is that they are compatible with the patient's immune system.…”

Large-Scale Production of Human iPSC-Derived Macrophages for Drug Screening

“…Recent studies successfully derived macrophage-like progenitor cells from human-induced Pluripotent Stem Cells (iPSCs). These cells can be further specialized, either by co-culturing with other tissue specific cell types or by simulating tissue environment with soluble factors, to cells that resemble key characteristics of tissue-resident macrophages (for simplicity, herein called “iPSC-derived macrophages”) [ 14 , 15 , 16 , 17 , 18 , 19 ]. In mouse embryonic development Myb-independent macrophages from the primitive streak populate most tissues before the start of definitive hematopoiesis [ 20 , 21 , 22 , 23 , 24 , 25 ] and renew in most tissues with only minor contributions from adult blood derived monocytes [ 26 , 27 ].…”

“…Interestingly, most of the iPSC-based differentiation protocols resemble Myb-independent primitive myelopoiesis, also shown for the one improved in this study by Buchrieser et al [ 28 ]. Hence, they are considered to be well suited for the generation of iPSC macrophages that model tissue resident macrophages of various tissues [ 7 , 29 ], displaying tissue-specific markers and functionality when compared to their primary counterparts [ 10 , 17 , 18 , 19 , 28 , 30 ]. Moreover, the method gives access to cells with a disease-relevant genetic background and readily allows genetic engineering (e.g., correction or introduction of a disease-causing mutation in the pluripotent state).…”