Tonsil-derived mesenchymal stem cells enhance allogeneic bone marrow engraftment via collagen IV degradation

Abstract

Background Co-transplantation of bone marrow cells (BMCs) and mesenchymal stem cells (MSCs) is used as a strategy to improve the outcomes of bone marrow transplantation. Tonsil-derived MSCs (TMSCs) are a promising source of MSCs for co-transplantation. Previous studies have shown that TMSCs or conditioned media from TMSCs (TMSC-CM) enhance BMC engraftment. However, the factors in TMSCs that promote better engraftment have not yet been identified. Methods Mice were subjected to a myeloablative regimen of busulfan and cyclophosphamide, and the mRNA expression in the bone marrow was analyzed using an extracellular matrix (ECM) and adhesion molecule-targeted polymerase chain reaction (PCR) array. Nano-liquid chromatography with tandem mass spectrometry, real-time quantitative PCR, western blots, and enzyme-linked immunosorbent assays were used to compare the expression levels of metalloproteinase 3 (MMP3) in MSCs derived from various tissues, including the tonsils, bone marrow, adipose tissue, and umbilical cord. Recipient mice were conditioned with busulfan and cyclophosphamide, and BMCs, either as a sole population or with control or MMP3-knockdown TMSCs, were co-transplanted into these mice. The effects of TMSC-expressed MMP3 were investigated. Additionally, Enzchek collagenase and Transwell migration assays were used to confirm that the collagenase activity of TMSC-expressed MMP3 enhanced BMC migration. Results Mice subjected to the myeloablative regimen exhibited increased mRNA expression of collagen type IV alpha 1/2 (Col4a1 and Col4a2). Among the various extracellular matrix-modulating proteins secreted by TMSCs, MMP3 was expressed at higher levels in TMSCs than in other MSCs. Mice co-transplanted with BMCs and control TMSCs exhibited a higher survival rate, weight recovery, and bone marrow cellularity compared with mice co-transplanted with BMCs and MMP3-knockdown TMSCs. Control TMSC-CM possessed higher collagenase activity against collagen IV than MMP3-knockdown TMSC-CM. TMSC-CM also accelerated BMC migration by degrading collagen IV in vitro. Conclusions Collectively, these results indicate that TMSCs enhance BMC engraftment by the secretion of MMP3 for the modulation of the bone marrow extracellular matrix.

How mesenchymal stem cell cotransplantation with hematopoietic stem cells can improve engraftment in animal models

“…In terms of HSCs, half of the selected studies[ 8 , 10 , 15 , 16 , 18 , 20 - 22 , 25 ] used cells from human umbilical cord blood (UCB), while the other half[ 9 , 11 - 14 , 17 , 18 , 23 , 24 ] used cells from animal BM (Table 1 ). Wu et al [ 20 ] reported the transplantation of the pool of human nucleated cells.…”

“…Huang et al [ 10 ] and Lim et al [ 21 ] reported the use of a mononuclear cell pool without any cell selection process, and the other 33.3% of the selected studies used human cells CD34+ before transplantation. The majority of studies[ 9 , 11 - 14 , 18 , 23 , 24 ] used the pool of nucleated cells (44.4%) from animals, with the exception of the study by Fernández-García et al [ 17 ] (5.6%) that used cells of lineage Sca-1+ cKit+ (LSK).…”

“…The transplantation process was reported using mainly 1 × 10 7 cells[ 9 - 11 , 21 ], ranging from 2.5 × 10 3 [ 8 ] to 1 × 10 7 cells, with exception of the study by Kornblit et al [ 23 ] that used 1.8 × 10 8 to 5.3 × 10 8 /kg cells in Beagle dogs with an administered volume of 50 mL. Four studies[ 8 , 10 , 15 , 25 ] compared two routes, intravenous (IV) versus intrabone (IB), but the majority of studies used the IV route (72.7%) with a volume of 100 to 250 µL, followed by 22.2% IB[ 8 , 10 , 15 , 25 ], being administered at a volume between 10 and 20 µL.…”

Procollagen C-Endopeptidase Enhancer 2 Secreted by Tonsil-Derived Mesenchymal Stem Cells Increases the Oxidative Burst of Promyelocytic HL-60 Cells

“…Because TMSCs successfully maintain MSC-specific properties during freezing and thawing, and also up to passage 15 during long-term in vitro cell culture [ 36 ], TMSCs are a good MSC candidate for clinical cell therapy. Our recent studies suggested that co-transplantation of bone marrow cells and TMSCs improved the outcomes of bone marrow transplantation by enhancing bone marrow cell engraftment [ 37 ] and thymus regeneration [ 38 ]. Because the infection is significantly associated with morbidity and mortality among patients receiving allogeneic bone marrow cells, enhancing neutrophil function is critical in maintaining host immune defences against invading microorganisms during bone marrow transplantation.…”

Conditioned medium from human tonsil-derived mesenchymal stem cells inhibits glucocorticoid-induced adipocyte differentiation

“…Our research group is currently investigating whether T-MSCs enhance bone marrow transplantation efficacy. In a series of experiments, we show that faster bone marrow reconstitution is accompanied by reduced bone marrow adiposity [ 35 , 36 ]. Further research identifying effector molecules that can enhance bone marrow reconstitution via inhibition of BMAT expansion is required.…”