Table 1 Main features and analysis performed in retrieved papers
Reference | Type of cells | Number of donors and age | Culture supplement | Type of HPL | Standard characterization (proliferation, immunophenotype, morphology) | Differentiation | Safety | Immunomodulatory/anti-inflammatory/angiogenic properties | |
|---|---|---|---|---|---|---|---|---|---|
FBS | HPL | ||||||||
Shanbhag et al. [11] | ASC BMSC | 3 8–14 yo | 10% | 5% | Home-made | Proliferation (DNA quantification) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin red S Staining, qPCR, ALP activity) | – | – |
Fuoco et al. [16] | ASC | 2 F 53–56 yo | 10% | 10% | Home-made | Proliferation (DT, MTT assay) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) Chondrogenic (Alcian Blue staining) | – | – |
Palombella et al. [5] | ASC | 3 m. 49 ± 2 yo | 10% | 5% | Heparin-free HPL, GMP Grade (Antibodies-online.com) | Proliferation (MTS assay) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) | – | Secreted cytokines (ELISA) Neurotrophic properties (qPCR, IF, co-culture with dorsal root ganglia) |
Gao et al. [17] | ASC | 8 40–80 yo | 10% | 5% 10% | Home-made | Proliferation (DT) Immunophenotype (FACS analysis) Pluripotency (qPCR, IF) Morphology (qualitative) | Osteogenic (ALP activity, Von Kossa staining and quantification) Chondrogenic (Alcian Blue staining and quantification) | – | – |
Becherucci et al. [18] | BMSC | 12 m. 25 yo | 10% | 5% | Home-made | Proliferation (PD) Immunophenotype (FACS analysis) Morphology (qualitative, FSC, SSC) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) Chondrogenic (Alcian Blue staining) | Relative telomere length | Mixed leukocyte reaction T-regulatory cell induction |
Boraldi et al. [19] | BMSC | 1 M 42 yo | 10% | 5% 8% | Stemulate, cook medical (+ / − heparin) Macopharma Lyset (+ heparin; Sclavo Diagnostic International) | Proliferation (PD) | – | – | – |
Pierce et al. [20] | BMSC (Lonza) | 3 | 10% | 10% | Home-made | Proliferation (DT, MTT assay, cell counting) Immunophenotype (FACS analysis) | – | – | Microarray gene expression analysis |
Fernandez-Rebollo et al. [21] | BMSC | 6 54–82 yo | 10% | 10% | Home-made | Proliferation (DT, PD, cell counting) Immunophenotype (FACS analysis) Morphology (qualitative, aspect ratio) Focal adhesions (IF) | Adipogenic (BODIPY/DAPI staining) Osteogenic (Alizarin Red staining) | Senescence (β-galactosidase staining) DNA-methylation analysis | Microarray gene expression analysis and semi-quantitative PCR |
Frese et al. [22] | ASC | 5 F m. 49 ± 8 yo | 10% | 10% | Home-made | Proliferation (DT) Immunophenotype (FACS analysis, IHC) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) Chondrogenic (Toluidine Blue staining) | – | – |
Juhl et al. [23] | BMSC | 1 M, 2 F 20–25 yo m. 22 yo | 10% | 5% | PLTMax (+ heparin) Stemulate, Cook Medical (+ / − heparin) | Proliferation (PD) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) Chondrogenic (Alcian Blue staining) | Genomic stability (Comparative Genomic Hybridization) | – |
Riis et al. [24] | ASC | 5 | 10% | 5% 10% | Stemulate, Cook Medical | Proliferation (DT, PD, CFU, cell counting) Immunophenotype (FACS analysis) Cell attachment on culture surface Morphology (qualitative, FSC, SSC) | – | – | – |
Castrèn et al. [25] | BMSC | 20–30 yo | 10% | 0.5% | Home-made | Immunophenotype (FACS analysis) | Osteogenic (ALP activity, Sirius Red staining and quantification, Alizarin Red staining and quantification, calcium content, qPCR) | – | – |
Hildner et al. [26] | ASC | 8 | 10% | 5% 10% | Home-made | Proliferation (PD) | Chondrogenic (Alcian Blue staining, IHC, GAG quantification, qPCR) | – | – |
Muraglia et al. [27] | BMSC | 3 | 10% | 0.5% 1% 2.5% | Home-made | Proliferation (PD, CFU, MTT assay) Immunophenotype (FACS analysis) | – | – | – |
Castiglia et al. [28] | BMSC | 19 0.5–39 yo | 10% | 10% | Home-made | Proliferation (PD, CFU) Immunophenotype (FACS analysis) Pluripotency (IF staining and quantification, qPCR) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Von Kossa staining) Chondrogenic (Alcian Blue staining) | Karyotype analysis Tumorigenesis test | – |
Fekete et al. [13] | BMSC | 3 | 5% 10% 20% | 5% 10% 20% | Home-made | Proliferation (cell counting, seeding density effect) | – | – | – |
Bernardi et al. [29] | BMSC | 4 | 10% | 2.5% 5% 7.5% 10% | Home-made | Proliferation (DT, PD) Immunophenotype (FACS analysis) | – | – | – |
Kinzebach et al. [12] | ASC BMSC | 3 F—ASC 3—BMSC | 2.5% 5% 7.5% 10% | 2.5% 5% 7.5% 10% | Home-made | Proliferation (ATP content assay, protein influence) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Von Kossa staining) | – | Mixed leukocyte reaction (only ASC) Secreted cytokines (ELISA) |
Menard et al. [30] | BMSC | nd | 10% | 8% | Home-made | Immunophenotype (FACS analysis) | – | – | Anti-inflammatory genes (qPCR) Inhibition of immune cell proliferation Mixed lymphocyte reaction IDO activity |
Trojahn Kølle et al. [31] | ASC | 4 F | 10% | 10% | Home-made | Proliferation (DT) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) Chondrogenic (Alcian Blue staining) | Chromosomal stability analysis | In vitro angiogenesis test Secreted cytokines (ELISA) |
Warnke et al. [32] | BMSC | 1 M 53 yo | 10% | 10% | Home-made | Proliferation (WST assay) Immunophenotype (FACS analysis) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) | – | – |
Azouna et al. [33] | BMSC | 13 16–41 yo m. 33 ± 2 yo | 10% | 5% 10% | Home-made | Proliferation (DT, CFU) Immunophenotype (FACS analysis) Morphology (qualitative) | Chondrogenic (Alcian Blue staining, qPCR, WB) | – | Secreted cytokines (ELISA) |
Gottipamula et al. [34] | BMSC | 4 | 10% | 10% | Home-made | Proliferation (DT, PD, CFU) Immunophenotype (FACS analysis) Morphology (qualitative) | Chondrogenic (Safranin O staining) | – | Mixed leukocyte reaction |
Cholewa et al. [35] | ASC | 5 | 10% | 10% | Home-made | Proliferation (PD, CFU, MTT assay) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining, qPCR) Osteogenic (Alizarin Red staining, qPCR) | Senescence (β-galactosidase staining) | – |
Flemming et al. [36] | BMSC | 5 21–72 yo | 10% | 10% | Home-made | Immunophenotype (FACS analysis) Morphology (qualitative) | – | – | Polyclonal stimulation Mixed leukocyte reaction Degranulation assay with T cells |
Castegnaro et al. [37] | ASC | 7 25–50 yo m. 40 yo | 10% | 10% | Home-made | Proliferation (PD, CFU) Immunophenotype (FACS analysis) | Adipogenic (Oil Red O staining, qPCR) Osteogenic (Von Kossa staining, qPCR) | ALDH activity | T-cells proliferation inhibition |
Chevallier et al. [38] | BMSC | 10 36–54 yo | 10% | 5% | Home-made | Proliferation (DT) | – | – | – |
Horn et al. [14] | BMSC | nd | 10% | 10% | Home-made | Proliferation (PD, CFU, MTT assay) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining, BODIPY/DAPI staining and quantification) Osteogenic (Alizarin Red staining) | Senescence (β-galactosidase staining) | – |
Schallmoser et al. [39] | BMSC | 1 F, 2Â M 9, 27, 36 yo | 10% | 10% | Home-made | Proliferation (PD, CFU) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) | Senescence (qPCR) | Microarray gene expression analysis |
Bieback et al. [40] | BMSC | 14 m. 22 yo | 10% | 10% | Home-made | Proliferation (DT, PD, CFU) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Von Kossa staining) | Telomerase activity | T-cells proliferation inhibition Secreted cytokines (cytokines antibody array) |
Blande et al. [15] | ASC | 9 FBS, 4 HPL | 10% | 10% | Home-made | Proliferation (DT) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) Osteogenic (Alizarin Red staining) | – | – |
Prins et al. [41] | BMSC | 9 4–74 yo | 10% | 5% | Home-made | Proliferation (PD, CFU) Immunophenotype (FACS analysis) Morphology (FSC, SSC) | Osteogenic (ALP expression) Chondrogenic (IF) | – | – |
Capelli et al. [42] | BMSC | 5 | 10% | 5% | Home-made | Proliferation (cell counting) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Sudan IV staining) Osteogenic (ALP expression) | – | Mixed leukocyte reaction PHA stimulation assay |
Schallmoser et al. [43] | BMSC | 2 F, 2 M 9–43 yo | 10% | 10% | Home-made | Proliferation (PD, CFU) Immunophenotype (FACS analysis) Morphology (qualitative) | Adipogenic (Oil Red O staining) | – | Secreted cytokines (multiplex detection) |
Doucet et al. [3] | BMSC | 10 | 10% | 5% | Home-made | Proliferation (cell counting, CFU) Immunophenotype (FACS analysis) | Osteogenic (ALP activity and immunohistochemistry, Von Kossa staining) | – | Mixed leukocyte reaction |