Hypertrophy is induced during the in vitrochondrogenic differentiation of human mesenchymal stem cells by bone morphogenetic protein-2 and bone morphogenetic protein-4 gene transfer

Table 1 Primer sequences and product sizes, for semi-quantitative and real-time RT-PCR

Gene	RT-PCR primer sequences (5'-3')	Annealing temp. (°C)	Product size (bp)	Cycles
Chondrogenic markers
COL II	Sense: TTTCCCAGGTCAAGATGGTC Antisense: CTTCAGCACCTGTC CACCA	58	374	35
AGC	Sense: TGAGGAGGGCTGGAACAAGTACC Antisense: GGAGGTGGTAATTGCAGGGAACA	54	392	30
COMP	Sense: CAGGACGACTTTGATGCAGA Antisense: AAGCTGGAGCTGTCTGGTA	54	312	32
FMD	Sense: CTTACCCCTATGGGGTGGAT Antisense: GTACATGGCCGTGAGGAAGT	54	389	35
SOX9	Sense: ATCTGAAGAAGGAGAGCGAG Antisense: TCAGAAGTCTCCAGAGCTTG	58	263	35
SOX9 (rt)	Sense: GGA GTGGAAGTTACTGACTGATG Antisense: AGGCGTTTTGCTTCGTCAATG	55	60
Hypertrophy and osteogenic markers
COL I	Sense: GGACACAATGGATTGCAAGG Antisense: TAACCACTGCTCCACTCTGG	54	461	32
COL X	Sense: CCCTTTTTGCTGCTAGTATCC Antisense: CTGTTGTCCAGGTTTTCCTGGCAC	54	468	25
OP	Sense: ACGCCGACCAAGGAAAACTC Antisense: GTCCATAAACCACACTATCACCTCG	51	483	35
ALP (rt)	Sense: TGGAGCTTCAGAAGCTCAACACCA Antisense: ATCTCGTTGTCTGAGTACCAGTCC	51	454
IHH	Sense: GAGGAGTCCCTGCATTATGA Antisense: CAGGAAAATGAGCACATCGC	54	321	30
RUNX2	Sense: ACAGATGATGACACTGCCACC Antisense: CATAGTAGAGATATGGAGTGCTGC	55	324	35
Internal control
EF1α	Sense: AGGTGATTATCCTGAACCATCC Antisense: AAAGGTGGATAGTCTGAGAAGC	54	234	25

rt: primer pairs, that have been used for real-time PCR only.
AGC = aggrecan core protein; ALP = alkaline phosphatase; COL = collagen; COMP = cartilage oligomeric matrix protein; EF1α = elongation factor 1α; FMD = fibromodulin; IHH = indian hedgehog; OP = osteopontin; RUNX2 = runt-related transcription factor 2; SOX9 = SRY (sex determining region Y) - box9.

ISSN: 1478-6362