Age-related Morphofunctional Changes in Sickle Cell Mice Bone Marrow Mesenchymal Stromal Cells.

BACKGROUND AND OBJECTIVES: Bone marrow mesenchymal stromal cells (BM-MSCs) are key elements of the hematopoietic niche and participate in the regulatory mechanisms of hematopoietic stem cells (HSCs). Hematological diseases can affect MSCs and their functions. However, the dysregulations caused by sickle cell disease (SCD) are not fully elucidated. This work explored changes in BM-MSCs and their relationship with age using sickle cell mice (Townes-SS). MATERIALS AND METHODS: BM-MSCs were isolated from Townes-SS, and control groups 30- and 60-day-old Townes-AA and C57BL/6 J. RESULTS: The BM-MSCs showed no morphological differences in culture and demonstrated a murine MSC-like immunophenotypic profile (Sca-1+, CD29+, CD44+, CD90.2+, CD31-, CD45-, and CD117-). Subsequently, all BM-MSCs were able to differentiate into adipocytes and osteocytes in vitro. Finally, 30-day-old BM-MSCs of Townes-SS showed higher expression of genes related to the maintenance of HSCs (Cxcl12, Vegfa, and Angpt1) and lower expression of pro-inflammatory genes (Tnfa and Il-6). However, 60-day-old BM-MSCs of Townes-SS started to show expression of genes related to reduced HSC maintenance and increased expression of pro-inflammatory genes. CONCLUSION: These results indicates age as a modifying factor of gene expression of BM-MSCs in the context of SCD.

Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation.

Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and regenerative medicine. In this article, we describe protocols to isolate MSCs from bone marrow (BM-MSCs) and adipose tissues (AT-MSCs), and methods to culture, characterize, and differentiate MSCs into osteoblasts, adipocytes, and chondrocytes. After the harvesting of cells from bone marrow by flushing the femoral diaphysis and enzymatic digestion of abdominal and inguinal adipose tissues, MSCs are selected by their adherence to the plastic tissue culture dish. Within 7 days, MSCs reach 70% confluence and are ready to be used in subsequent experiments. The protocols described here are easy to perform, cost-efficient, require minimal time, and yield a cell population rich in MSCs.

Adipogenic differentiation of murine bone marrow mesenchymal stem cells induced by visible light via photo- induced biomodulation.

BACKGROUND: Bone marrow mesenchymal stem cells (BM-MSCs) are undifferentiated cells that can proliferate and differentiate into specialized cells for tissue self-repair. Low-level laser (LLL) can induce biomodulatory effects such as cellular proliferation, differentiation, and migration. We investigated the biomodulatory effects of the photoactive compound chloroaluminum phthalocyanine nanoemulsion (AlClPc/NE) on the adipogenic differentiation of BM-MSCs, when combined with LLL (AlClPc/NE-LLL). METHODS: The BM-MSCs used in this work were isolated from green fluorescent protein-positive (GFP+) C57BL6 mice. Cells were first treated with AlClPc/NE, a well-designed photoactive nano-drug and were then subjected to in vitro expansion, morphological and immunophenotypic characterization, and cellular cytotoxicity analysis. Subsequently, BM-MSCs were induced to differentiate into adipocytes by photo-induced biomodulation with AlClPc/NE-LLL. RESULTS: Our results showed that the isolated cell population was consistent with murine BM-MSCs. The cellular cytotoxicity analysis revealed that the optimal nanoemulsion dose to induce BM-MSC biomodulation was 5.0 µmol/L. Twenty-four hours following treatment with AlClPc/NE, BM-MSC were subjected to visible light irradiation of 20 mJ/cm2 at 670 nm. Six days after photo-induced biomodulation, cells maintained high GFP expression level, and expressed detectable mRNA levels of adipogenic genes (lipoprotein lipase and PPARγ); formation of lipid vacuoles was observed, and the cells did not show any tumorigenic potential in vivo. CONCLUSIONS: Our results indicated that photo-induced biomodulation via visible light using AlClPc/NE and LLL can induce adipogenic differentiation of murine BM-MSCs. Therefore, cell therapy with BM-MSCs and photo-induced biomodulation may contribute to the development of new therapeutic strategies that are faster and more effective than traditional methods to trigger MSC differentiation.

Effect of cell therapy with osteoblasts differentiated from bone marrow or adipose tissue stromal cells on bone repair.

Aim: The aim of this study was to investigate the effect of local injection of osteoblasts differentiated from bone marrow (BM-OB) or adipose tissue (AT-OB) mesenchymal stromal cells on bone tissue formation. Materials & methods: Defects were created in rat calvaria and injected with BM-OB or AT-OB and phosphate-buffered saline without cells were injected as control. Bone formation was evaluated 4 weeks postinjection. Results: Injection of BM-OB or AT-OB resulted in higher bone formation than that obtained with control. The bone tissue induced by cell injections exhibited similar mechanical properties as those of pristine calvarial bone, and its molecular cues suggested the occurrence of a remodeling process. Conclusion: Results of this study demonstrated that cell therapy with osteoblasts induced significant bone formation that exhibited the same quality as that of pre-existent bone.