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Peer-reviewedCC BY-NCInternational Journal of Molecular and Cellular Medicine

Biochemical profiling of adipose-derived mesenchymal stem cell conditioned media: Reproducible protein quantification and growth factor analysis

  1. 1Department of Medical Laser, Medical Laser Research Center, Yara Institute, ACECR, Tehran, Iran.
  2. 2Department of Photodynamic, Medical Laser Research Center, Yara Institute, ACECER, Tehran. Iran

Abstract

Mesenchymal stem cell-conditioned media (MSC-CM) represents a promising cell-free therapeutic strategy in regenerative medicine, leveraging the paracrine effects of stem cells. Standardized biochemical characterization of MSC-CM is crucial for ensuring its reproducibility and therapeutic potential. However, statistical validation of batch-to-batch reproducibility has been rarely reported. Therefore, this study aimed to establish a reproducible protocol for AD-MSC-CM production and quantitatively demonstrate batch-to-batch reproducibility of total protein and key growth factors. Adipose-derived MSCs (AD-MSCs) were isolated from rat adipose tissue, expanded, and used to produce conditioned media (MSC-CM). The MSC-CM was biochemically profiled using the Bradford assay for total protein concentration and specific enzyme-linked immunosorbent assays (ELISAs) for quantifying Transforming Growth Factor-Beta (TGF-β) and Vascular Endothelial Growth Factor (VEGF). Analyses were performed on three independent biological replicates. The MSC-CM showed a significant and consistent increase in total protein concentration compared to control serum-free medium. Quantitative analysis revealed reproducible and stable secretion of key growth factors. The concentrations of both TGF-β and VEGF showed no significant variation across the biological replicates, confirming the high reproducibility of the MSC-CM preparation protocol. This study established a robust method for generating reproducible AD-MSC-CM, characterized by a stable and enriched content of total protein and specific growth factors (TGF-β and VEGF). The findings confirm the reliability of the production process and highlight the hypothesized potential of this well-characterized MSC-CM as a consistent therapeutic agent for applications requiring immunomodulation and angiogenesis, pending direct functional validation.

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BibTeX
@article{nasiri2026biochemical,
  title = {{Biochemical profiling of adipose-derived mesenchymal stem cell conditioned media: Reproducible protein quantification and growth factor analysis}},
  author = {Nahid Nasiri and Shaghayegh Gholami and Armin Nazemi Zadeh and Seyed Mehdi Tabaei},
  journal = {International Journal of Molecular and Cellular Medicine},
  year = {2026},
  month = jun,
  note = {CC BY-NC},
}
RIS
TY  - JOUR
TI  - Biochemical profiling of adipose-derived mesenchymal stem cell conditioned media: Reproducible protein quantification and growth factor analysis
AU  - Nahid Nasiri
AU  - Shaghayegh Gholami
AU  - Armin Nazemi Zadeh
AU  - Seyed Mehdi Tabaei
PY  - 2026
DA  - 2026/06/01
JO  - International Journal of Molecular and Cellular Medicine
AB  - Mesenchymal stem cell-conditioned media (MSC-CM) represents a promising cell-free therapeutic strategy in regenerative medicine, leveraging the paracrine effects of stem cells. Standardized biochemical characterization of MSC-CM is crucial for ensuring its reproducibility and therapeutic potential. However, statistical validation of batch-to-batch reproducibility has been rarely reported. Therefore, this study aimed to establish a reproducible protocol for AD-MSC-CM production and quantitatively demonstrate batch-to-batch reproducibility of total protein and key growth factors. Adipose-derived MSCs (AD-MSCs) were isolated from rat adipose tissue, expanded, and used to produce conditioned media (MSC-CM). The MSC-CM was biochemically profiled using the Bradford assay for total protein concentration and specific enzyme-linked immunosorbent assays (ELISAs) for quantifying Transforming Growth Factor-Beta (TGF-β) and Vascular Endothelial Growth Factor (VEGF). Analyses were performed on three independent biological replicates. The MSC-CM showed a significant and consistent increase in total protein concentration compared to control serum-free medium. Quantitative analysis revealed reproducible and stable secretion of key growth factors. The concentrations of both TGF-β and VEGF showed no significant variation across the biological replicates, confirming the high reproducibility of the MSC-CM preparation protocol. This study established a robust method for generating reproducible AD-MSC-CM, characterized by a stable and enriched content of total protein and specific growth factors (TGF-β and VEGF). The findings confirm the reliability of the production process and highlight the hypothesized potential of this well-characterized MSC-CM as a consistent therapeutic agent for applications requiring immunomodulation and angiogenesis, pending direct functional validation.
SN  - 2251-9637
SN  - 2251-9645
ER  -