Adipose-derived mesenchymal stem cells: Source-dependent heterogeneity, translational challenges, and emerging precision strategies

Table 1 Source-dependent factors influencing the biological characteristics of adipose-derived mesenchymal stem cells

Factor	Representative condition/source	Main affected properties	Representative findings	Ref.
Donor age	Young vs aged donors	Proliferation, senescence, osteogenesis	Aging is associated with reduced proliferation, earlier senescence, and impaired osteogenesis	[8-10,33,34]
Metabolic status	Obesity	Inflammatory phenotype, mitochondrial function, secretome	Obesity is associated with inflammatory activation and reduced regenerative capacity	[5,37,38,40]
Disease background	Type 2 diabetes mellitus	Migration, angiogenesis, senescence	Diabetic ADSCs show impaired migration and angiogenic support and increased senescence susceptibility	[37,39,41]
Sex	Female vs male donors	Immunomodulation, transcriptomic profile	Sex-related differences may influence immunomodulatory and molecular features	[12,13,32]
Adipose tissue depot	Subcutaneous vs visceral fat	Molecular profile, osteogenic/adipogenic tendency	Visceral ADSCs show a more inflammatory profile, whereas subcutaneous ADSCs may favor osteogenesis	[1,3,4,37]
Subcutaneous regional depot	Abdominal vs gluteal/thigh depots	Proliferation, differentiation potential	Different subcutaneous depots are not functionally equivalent	[1,5]
Isolation method	Enzymatic digestion	Yield, phenotype	Higher cell yield, but phenotype may be affected by digestion conditions	[37]
Isolation method	Mechanical separation	Heterogeneity preservation	Better preservation of native subpopulations, but lower yield	[37]
Culture conditions	Hypoxia	Metabolism, stemness, secretome	Hypoxia may enhance stemness and paracrine activity	[14,15]
Culture conditions	2D vs 3D culture	Functional state, secretome potency	3D culture may improve regenerative secretome features	[14,15]
Epigenetic regulation	DNA methylation/tissue memory	Molecular memory, differentiation	Epigenetic regulation contributes to source-dependent heterogeneity	[30,43]
Secretome heterogeneity	Healthy vs diseased source	Cytokines, growth factors, EV composition	Diseased sources tend to generate more pro-inflammatory secretomes	[15,37,45]

ADSCs: Adipose-derived mesenchymal stem cells; 2D: Two-dimensional; 3D: Three-dimensional; EV: Extracellular vesicle.

Table 2 Major translational challenges in adipose-derived mesenchymal stem cell-based therapy and potential strategies to address them

Challenge	Main problem	Potential strategy	Ref.
Donor heterogeneity	Major variability in phenotype and function	Source-aware donor screening	[5,6,8,12,37]
Depot-specific variability	Different depots yield distinct ADSCs	Match source to therapeutic indication	[1,3,4,37]
Regional variability	Subcutaneous depots are not equivalent	Improve anatomical precision in tissue collection	[1,5]
Isolation inconsistency	Isolation methods alter downstream cell behavior	Standardize processing workflows	[37]
Culture-induced drift	Serum, oxygen, and culture format reshape phenotype	Use defined media and standardized culture systems	[6,14,15,29]
Cryopreservation variability	Freezing and thawing affect cell quality	Optimize banking and post-thaw validation	[30,46]
Lack of potency assays	Functional release criteria remain unclear	Develop source-aware potency markers	[6,30,47,50]
Incomplete molecular stratification	Bulk analyses miss functional subpopulations	Apply single-cell and multi-omics profiling	[52-55]
Secretome heterogeneity	Secretome products are highly source- and process-dependent	Standardize production and characterization	[56-60]
Manufacturing inconsistency	Scale-up and reproducibility remain limited	Introduce source-aware manufacturing and QC systems	[54,63,65]
Limited trial stratification	Clinical trials often insufficiently stratify source/product features	Incorporate source-related variables into trial design	[50,52,61,62,64]

ADSCs: Adipose-derived mesenchymal stem cells; QC: Quality control.