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Table 2 Analyses in chondrogenically differentiated aggregate cultures of human mesenchymal stem cells (hMSCs)

From: SOX9 gene transfer via safe, stable, replication-defective recombinant adeno-associated virus vectors as a novel, powerful tool to enhance the chondrogenic potential of human mesenchymal stem cells

Assay

No vector

rAAV-lacZ

rAAV-FLAG-hSOX9

 

Day 7

Day 21

Day 7

Day 21

Day 7

Day 21

Diameter (μm)

632 (6)

618 (4)c

630 (2)

614 (3)c

729 (4)a, b

982 (5)a, b, c

Cell density (cells/mm2)

433 (7)

424 (2)c

438 (4)

426 (1)c

437 (4)

425 (5)c

WST-1 (OD450 nm)

1.33 (0.17)

0.62 (0.12)c

1.29 (0.21)

0.67 (0.12)c

1.31 (0.15)

0.65 (0.14)c

DNA (ng/mg total proteins)

0.92 (0.02)

0.87 (0.02)c

0.90 (0.01)

0.86 (0.01)c

0.92 (0.01)

0.88 (0.02)c

Toluidine blue intensity (pixels)

ND

167 (4)

ND

169 (5)

ND

914 (7)a, b

Proteoglycans (ng/mg total proteins)

0.61 (0.02)

0.79 (0.04)c

0.59 (0.03)

0.80 (0.02)c

3.37 (0.06)a, b

4.68 (0.07)a, b, c

Proteoglycans/DNA (ng/ng)

0.67 (0.04)

0.91 (0.03)c

0.66 (0.03)

0.93 (0.02)c

3.67 (0.04)a, b

5.32 (0.08)a, b, c

Type II collagen intensity (%)

ND

51 (3)

ND

49 (2)

ND

85 (3)a, b

Type II collagen (pg/mg total proteins)

13 (2)

18 (3)c

14 (2)

19 (3)c

75 (4)a, b

112 (3)a, b, c

Type II collagen/DNA (pg/ng)

14 (2)

21 (2)c

15 (1)

22 (3)c

81 (4)a, b

127 (8)a, b, c

Type I collagen intensity (%)

ND

44 (2)

ND

42 (4)

ND

6 (2)a, b

Type X collagen intensity (%)

ND

45 (3)

ND

46 (2)

ND

8 (2)a, b

Alizarin red intensity (pixels)

ND

57 (3)

ND

56 (2)

ND

5 (2)a, b

ALP activity (pg/mg total proteins)

ND

57 (2)

ND

55 (1)

ND

25 (1)a, b

ALP/DNA (pg/ng)

ND

66 (2)

ND

64 (3)

ND

28 (1)a, b

  1. Values are given as mean (SD) with a vector dose of 100 μl. ALP, alkaline phosphatase; ND, not done. Statistically significant compared with acondition without vector treatment, brAAV-lacZ, and cearlier time point.