Table 1.
Induction for the Differentiation of Mesenchymal Lineages and the Detection of Specific Markers Using Histochemical Staining, IHC, and RT-PCR
| Lineage, staining, and gene | Direction | Primer sequence |
|---|---|---|
| Adipogenic lineage (staining: HCS LipidTOX neutral lipid stain, peroxisome proliferation activated receptor γ (PPARγ) | ||
| PPARG | Sense | 5′-GCTCCGTGGATCTCTCCGTAATG-3′ |
| Antisense | 5′-AATTGCCATGAGGGAGTTGGAAGG-3′ | |
| CEBPA | Sense | 5′-TCGACATCAGCGCCTACATC-3′ |
| Antisense | 5′-CTTGTCCACCGACTTCTTGG-3′ | |
| LPL | Sense | 5′-CAAAACTTGTGGCCGCCCTGTA-3′ |
| Antisense | 5′-GGGGACCCTCTGGTGAATGTGTGT-3′ | |
| Myogenic lineage [staining: alpha smooth muscle actin (α-SMA)] | ||
| ACTA2 | Sense | 5′-CCGGGAGAAAATGACTCAAA-3′ |
| Antisense | 5′-GCGTCCAGAGGCATAGAGAG-3′ | |
| CNN1 | Sense | 5′-CGTCGCATCGGCAACAACTTCAT-3′ |
| Antisense | 5′-ACCTTGTTTCCTTTCGTCTTCGC-3′ | |
| CALD1 | Sense | 5′-ACAGTCACCAAGTCCTACCAGAAGAATG-3′ |
| Antisense | 5′-CCTCCAGGGCGGCTGAAAGT-3′ | |
| Osteogenic lineage (staining: alkaline phosphatase osteopontin) | ||
| ATHS | Sense | 5′-TGGAGCTTCAGAAGCTCAACACCA-3′ |
| Antisense | 5′-ATCTCGTTGTCTGAGTACCAGTCC-3′ | |
| RUNX2 | Sense | 5′-CTCACTACCACACCTACCTGCCAC-3′ |
| Antisense | 5′-TCAATATGGTCGCCAAACAGATTC-3′ | |
| SPP1 | Sense | 5′-AGGAGGAGGCAGAGCACA-3′ |
| Antisense | 5′-CTGGTATGGCACAGGTGATG-3′ | |
| PTH1R | Sense | 5′-CCTCACCGTAGCTGTGCTCATCCT-3′ |
| Antisense | 5′-GCCCCTCCACCAGAATCCAGTAG-3′ | |
| Chondrogenic lineage (staining: 1% Alcian Blue, Safranin O, collagen type II) | ||
| COL2A1 | Sense | 5′-CACTCCTGGCACTGATGGTCCC-3′ |
| Antisense | 5′-CTTCTCCCTTCTCGCCGTTAGCAC-3′ | |
| COL10A | Sense | 5′-GGACACAATGGAGAGGCTGGC-3′ |
| Antisense | 5′-ATGACTGCTTGACCTGGTGGGC-3′ | |
| GAPDH | Sense | 5′-ACCACAGTCCATGCCATCAC-3′ |
| Antisense | 5′-TCCACCACCCTGTTGCTGTA-3′ | |
ACTA2 (encodes α-SMA protein): actin, alpha 2, smooth muscle; ATHS (previously ALPL): alkaline phosphatase: atherosclerosis susceptibility (lipoprotein associated); CALD1: caldesmon 1; CEBPA: CCAAT/enhancer binding protein (C/EBP), alpha; CNN1: calponin 1, basic, smooth muscle; ATHS: COL10A1: collagen, type X, alpha 1; COL2A1: collagen, type II, alpha 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; LPL: lipoprotein lipase; PPARG: peroxisome proliferator-activated receptor gamma; PTH1R: parathyroid hormone 1 receptor; RUNX2: runt-related transcription factor 2; SPP1 (previously OPN, osteopontin): secreted phosphoprotein 1.