Table 2 Culture media formulations used for differentiation assays
Medium | Basal media | Serum | Coating/coating-free supplements | Supplementation |
|---|---|---|---|---|
Adipogenic (FBS) | DMEM/F-12 | 10% Fetal bovine serum | None | 1% GlutaMAX, 1% p/s, 33 μM biotin (Sigma), 1 μM dexamethasone (Sigma), 100 nM insulin (Life Technologies), 17 μM pantothenate (Fluka, Buchs, Switzerland), 250 μM isobutylmethylxanthine (IBMX; Sigma) for 48-hour induction after cell seeding |
Osteogenic (FBS) | DMEM/F-12 | 10% Fetal bovine serum | None | 1% GlutaMAX, 1% p/s, 150 μM L-ascorbic acid 2-phosphate (Sigma), 10 mM β-glycerophosphate (Sigma), 10 nM dexamethasone (Sigma) |
Chondrogenic (FBS/HS) | DMEM/F-12 | None | None | 1% GlutaMAX, 0.3% p/s, 10 mg/ml human serum albumin (Sigma), 8 μg/ml holo-transferrin human (Sigma), 5 ng/ml sodium selenite (Sigma), 10 μg/ml insulin (Life Technologies), 1 μg/ml linoleic acid (Sigma), 50 μM L-ascorbic acid 2-phosphate (Sigma), 55 μM sodium pyruvate (Life Technologies), 23 μM L-proline (Sigma), 10 ng/ml TGF-β (Sigma) |
Adipogenic (HS) | DMEM/F-12 | 10% Human serum | None | 1% GlutaMAX, 1% p/s, 33 μM biotin, 1 μM dexamethasone, 100 nM insulin, 17 μM pantothenate, 250 μM IBMX for 48-hour induction after cell seeding |
Osteogenic (HS) | DMEM/F-12 | 10% Human serum | None | 1% GlutaMAX, 1% p/s, 150 μM L-ascorbic acid 2-phosphate, 10 mM β-glycerophosphate, 10 nM dexamethasone |
Adipogenic (XF/SF CS) | StemPro MSC | none | CELLstart™ coating | StemPro MSC SFM XenoFree supplement, 1% GlutaMAX, 0.3% p/s, 33 μM biotin, 1 μM dexamethasone, 100 nM insulin, 17 μM pantothenate, 250 μM IBMX for 48-hour induction after cell seeding |
SFM | ||||
Osteogenic (XF/SF CS) | StemPro MSC | none | CELLstart™ coating | StemPro MSC SFM XenoFree supplement, 1% GlutaMAX, 0.3% p/s, 150 μM L-ascorbic acid 2-phosphate, 10 mM β-glycerophosphate, 10 nM dexamethasone |
SFM | ||||
Chondrogenic (XF/SF CS and XF/SF CM) | StemPro MSC | None | None | 1% GlutaMAX, 0.3% p/s, 10 mg/ml human serum albumin, 8 μg/ml holo-transferrin human, 5 ng/ml sodium selenite, 10 μg/ml insulin, 1 μg/ml linoleic acid, 50 μM L-ascorbic acid 2-phosphate, 55 μM sodium pyruvate, 23 μM L-proline, 10 ng/ml TGF-β |
SFM | ||||
Adipogenic (XF/SF CM) | StemPro MSC | None | Coating Matrix Kit | StemPro MSC SFM XenoFree supplement, 1% GlutaMAX, 0.3% p/s, 33 μM biotin, 1 μM dexamethasone, 100 nM insulin, 17 μM pantothenate, 250 μM IBMX for 48-hour induction after cell seeding |
SFM | ||||
Osteogenic (XF/SF CM) | StemPro MSC | None | Coating Matrix Kit | StemPro MSC SFM XenoFree supplement, 1% GlutaMAX, 0.3% p/s, 150 μM L-ascorbic acid 2-phosphate, 10 mM beta-glycerophosphate, 10 nM dexamethasone |
SFM |