Development of a two-step protocol for culture expansion of human annulus fibrosus cells with TGF-β1 and FGF-2

Table 1 Primers used in the real-time polymerase chain reaction analysis

Target gene	Forward primer	Reverse primer	GenBank accession no.	Roche probe no.
Housekeeping gene
GAPDH	agccacatcgctcagacac	gcccaatacgaccaaatcc	NM_002046	#60
Anabolic genes
COL IA1	gggattccctggacctaaag	ggaacacctcgctctcc	NM_000088	#67
COL IIIA1	ctggaccccagggtcttc	catctgatccagggtttcca	NM_000090	#20
Aggrecan	cctccccttcacgtgtaaaa	gctccgcttctgtagtctgc	NM_01135	#76
Catabolic genes
MMP-1	gctaacctttgatgctataactacga	tttgtgcgcatgtagaatctg	NM_001145938.1	#7
MMP-3	caaaacatatttctttgtagaggacaa	ttcagctatttgcttgggaaa	NM_002422	#36
ADAMTS-4	ccaggcactgggctactact	gaacagggggtcccatcta	NM_005099	#67
AF putative genes
COL1A1	gggattccctggacctaaag	ggaacacctcgctctcc	NM_000088	#67
COL2A1	tggtgctaatggcgagaag	cccagtctctccacgttcac	NM_033150.2	#4
COL5A1	gggattccctggacctaaag	ggaacacctcgctctcc	NM_000093.3	#42
COL12A1	cctggatgaggaggtgtttg	cagcactggcgacttagaaa	NM_004370.5	#89
SFRP-2	gctagcagcgaccacctc	tttttgcaggcttcacatacc	NM_003013.2	#83

ADAMTS a disintegrin and metallopeptidase with thrombospondin motif, COL collagen, GAPDH glyceraldehyde-3-phosphate dehydrogenase, MMP matrix metallopeptidase, SFRP secreted frizzled-related protein

ISSN: 1757-6512