Proposal of Simplified Standardization of the Cell-Growth-Promoting Activity of Human Adipose Tissue Mesenchymal Stromal Cell Culture Supernatants
Abstract
:1. Introduction
2. Results
2.1. Construction of Protocols for CM Production and Cell-Proliferation Assay
2.2. Cell-Proliferation-Promoting Activities of CMs
2.3. Cell-Proliferation-Promoting Activities of CM after 1 Year of Refrigerated Storage
2.4. Cell-Proliferation-Promoting Activities of CM with Young and Aged Cells
2.5. Cell-Proliferation-Promoting Activities of CM Fractionated by Molecular Weight
3. Discussion
4. Materials and Methods
4.1. Production of CMs
4.2. Assay of Cell-Proliferation-Promoting Activities
4.3. Preparation of Aged Cells, CM Fractionation, and Glucose Determination
4.4. Statistics
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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#PT-5006, Human adipose derived stem cells (1) | KW-4209, Human pre-adipocytes (1) | |||
Source (1) | Human adipose tissue obtained as lipoaspirate from patients undergoing elective liposuction surgery procedures. | Adult adipose tissue | ||
Passage number at shipment (1) | 1 | 2 | ||
Cell viability, % (2) | 88.0 ± 9.4, n = 5 | 88.9 ± 4.8, n = 12 | ||
Viable cell number (×106) per tube (2) | 1.26 ± 0.24, n = 5 | 0.99 ± 0.28, n = 12 | ||
Cellular changes in CM production process | ||||
Initial cell density (/cm2) | 7318 ± 99, n = 2 (3) | 6310 ± 310, n = 4 | ||
Medium for CM (4) | FBS- | FBS+ | FBS-, n = 2 | FBS+, n = 2 |
Cell viability, % | 96.2 | 97.9 | 95.1 ± 3.8 | 93.1 ± 6.1 |
Cell density (/cm2) at the time of harvest | 12,583 | 15,232 | 20,861 ± 728 | 27,881 ± 1788 |
Fold increase (5) | 1.70 | 2.11 | 3.40 ± 0.26 | 4.32 ± 0.38 |
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Enosawa, S.; Kobayashi, S.; Kobayashi, E. Proposal of Simplified Standardization of the Cell-Growth-Promoting Activity of Human Adipose Tissue Mesenchymal Stromal Cell Culture Supernatants. Int. J. Mol. Sci. 2024, 25, 5197. https://doi.org/10.3390/ijms25105197
Enosawa S, Kobayashi S, Kobayashi E. Proposal of Simplified Standardization of the Cell-Growth-Promoting Activity of Human Adipose Tissue Mesenchymal Stromal Cell Culture Supernatants. International Journal of Molecular Sciences. 2024; 25(10):5197. https://doi.org/10.3390/ijms25105197
Chicago/Turabian StyleEnosawa, Shin, Sho Kobayashi, and Eiji Kobayashi. 2024. "Proposal of Simplified Standardization of the Cell-Growth-Promoting Activity of Human Adipose Tissue Mesenchymal Stromal Cell Culture Supernatants" International Journal of Molecular Sciences 25, no. 10: 5197. https://doi.org/10.3390/ijms25105197