Bone Marrow Microenvironment in Light-Chain Amyloidosis: In Vitro Expansion and Characterization of Mesenchymal Stromal Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Enrollment of Patients and Healthy Donors
2.2. AL-MSCs Isolation and Expansion
2.3. Characterization of Ex Vivo Expanded AL-MSCs
2.3.1. Flow Cytometry
2.3.2. Differentiation Assay
2.3.3. Proliferative Capacity
2.3.4. Senescence Assay
2.3.5. Gene Expression Profile
2.4. ALMC-2 Cell Line
2.5. Co-Culture of AL-MSCs, HD-MSCs, and ALMC-2
2.6. Evaluation of λL Chain Levels in Culture Supernatants
2.7. Quantification of IgG and IL-6 Levels in Culture Supernatants
2.8. Intracellular λLight Chains
2.9. Co-Culture of AL-MSCs, HD-MSCs, and Peripheral Blood Mononuclear Cells (PBMCs)
2.10. Statistical Analysis
3. Results
3.1. Characterization of Ex Vivo Expanded AL-MSCs
Expression of Growth Factor Receptors in MSCs
3.2. Modulatory Effect of MSCs Co-Cultured with ALMC-2 Cells
3.3. Modulatory Effect of MSCs Co-Cultured with HD-PBMCs
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Subjects | n (%) | ||
Patients | 57 | ||
Male sex | 33 (57.9) | ||
Median (IQR) | |||
Age, yrs | 67 (60–73) | ||
% BMPC * | 9 (6–13) | ||
dFLC, mg/L ** | 175.3 (79.1–444.8) | ||
Organ | n (%) | ||
Heart | 43 (75.4) | ||
Kidney | 27 (47.4) | ||
Soft tissues | 13 (22.8) | ||
Liver | 3 (5.3) | ||
M protein | |||
Type | (n) | Type | (n) |
IgAκ | 1 | IgAλ | 7 |
IgGκ | 4 | IgGλ | 12 |
IgMκ | 0 | IgMλ | 3 |
κ | 9 | λ | 21 |
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Valsecchi, C.; Croce, S.; Maltese, A.; Montagna, L.; Lenta, E.; Nevone, A.; Girelli, M.; Milani, P.; Bosoni, T.; Massa, M.; et al. Bone Marrow Microenvironment in Light-Chain Amyloidosis: In Vitro Expansion and Characterization of Mesenchymal Stromal Cells. Biomedicines 2021, 9, 1523. https://doi.org/10.3390/biomedicines9111523
Valsecchi C, Croce S, Maltese A, Montagna L, Lenta E, Nevone A, Girelli M, Milani P, Bosoni T, Massa M, et al. Bone Marrow Microenvironment in Light-Chain Amyloidosis: In Vitro Expansion and Characterization of Mesenchymal Stromal Cells. Biomedicines. 2021; 9(11):1523. https://doi.org/10.3390/biomedicines9111523
Chicago/Turabian StyleValsecchi, Chiara, Stefania Croce, Alice Maltese, Lorenza Montagna, Elisa Lenta, Alice Nevone, Maria Girelli, Paolo Milani, Tiziana Bosoni, Margherita Massa, and et al. 2021. "Bone Marrow Microenvironment in Light-Chain Amyloidosis: In Vitro Expansion and Characterization of Mesenchymal Stromal Cells" Biomedicines 9, no. 11: 1523. https://doi.org/10.3390/biomedicines9111523