Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro

Chua Kien Hui, F. Raduan, W. K Z Wan Safwani, N. F M Manzor, B. Pingguan-Murphy, S. Sathapan

Research output: Contribution to journalArticle

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Abstract

Objectives: This study investigated effects of reduced serum condition and vascular endothelial growth factor (VEGF) on angiogenic potential of adipose stromal cells (ASCs) in vitro. Materials and methods: Adipose stromal cells were cultured in three different types of medium: (i) F12/DMEM (FD) supplemented with 10% FBS from passage 0 (P0) to P6; (ii) FD supplemented with 2% FBS at P6; and (iii) FD supplemented with 2% FBS plus 50 ng/ml of VEGF at P6. Morphological changes and growth rate of ASCs were recorded. Changes in stemness, angiogenic and endogenic genes' expressions were analysed using Real-Time PCR. Results: Adipose stromal cells changed from fibroblast-like shape when cultured in 10% FBS medium to polygonal when cultured in 2% FBS plus VEGF-supplemented medium. Their growth rate was lower in 2% FBS medium, but increased with addition of VEGF. Real-Time PCR showed that ASCs maintained most of their stemness and angiogenic genes' expression in 10% FBS at P1, P5 and P6, but this increased significantly in 2% FBS at P6. Endogenic genes expression such as PECAM-1, VE chaderin and VEGFR-2 decreased after serial passage in 10% FBS, but increased significantly at P6 in 2% FBS. Addition of VEGF did not cause any significant change in gene expression level. Conclusion: Adipose stromal cells had greater angiogenic potential when cultured in reduced serum conditions. VEGF did not enhance their angiogenic potential in 2% FBS-supplemented medium.

Original languageEnglish
Pages (from-to)300-311
Number of pages12
JournalCell Proliferation
Volume46
Issue number3
DOIs
Publication statusPublished - Jun 2013

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Stromal Cells
Vascular Endothelial Growth Factor A
Serum
Gene Expression
Real-Time Polymerase Chain Reaction
CD31 Antigens
Serial Passage
Vascular Endothelial Growth Factor Receptor-2
Growth
In Vitro Techniques
Fibroblasts

ASJC Scopus subject areas

  • Cell Biology

Cite this

Kien Hui, C., Raduan, F., Wan Safwani, W. K. Z., Manzor, N. F. M., Pingguan-Murphy, B., & Sathapan, S. (2013). Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro. Cell Proliferation, 46(3), 300-311. https://doi.org/10.1111/cpr.12029

Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro. / Kien Hui, Chua; Raduan, F.; Wan Safwani, W. K Z; Manzor, N. F M; Pingguan-Murphy, B.; Sathapan, S.

In: Cell Proliferation, Vol. 46, No. 3, 06.2013, p. 300-311.

Research output: Contribution to journalArticle

Kien Hui, C, Raduan, F, Wan Safwani, WKZ, Manzor, NFM, Pingguan-Murphy, B & Sathapan, S 2013, 'Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro', Cell Proliferation, vol. 46, no. 3, pp. 300-311. https://doi.org/10.1111/cpr.12029
Kien Hui, Chua ; Raduan, F. ; Wan Safwani, W. K Z ; Manzor, N. F M ; Pingguan-Murphy, B. ; Sathapan, S. / Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro. In: Cell Proliferation. 2013 ; Vol. 46, No. 3. pp. 300-311.
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AU - Pingguan-Murphy, B.

AU - Sathapan, S.

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N2 - Objectives: This study investigated effects of reduced serum condition and vascular endothelial growth factor (VEGF) on angiogenic potential of adipose stromal cells (ASCs) in vitro. Materials and methods: Adipose stromal cells were cultured in three different types of medium: (i) F12/DMEM (FD) supplemented with 10% FBS from passage 0 (P0) to P6; (ii) FD supplemented with 2% FBS at P6; and (iii) FD supplemented with 2% FBS plus 50 ng/ml of VEGF at P6. Morphological changes and growth rate of ASCs were recorded. Changes in stemness, angiogenic and endogenic genes' expressions were analysed using Real-Time PCR. Results: Adipose stromal cells changed from fibroblast-like shape when cultured in 10% FBS medium to polygonal when cultured in 2% FBS plus VEGF-supplemented medium. Their growth rate was lower in 2% FBS medium, but increased with addition of VEGF. Real-Time PCR showed that ASCs maintained most of their stemness and angiogenic genes' expression in 10% FBS at P1, P5 and P6, but this increased significantly in 2% FBS at P6. Endogenic genes expression such as PECAM-1, VE chaderin and VEGFR-2 decreased after serial passage in 10% FBS, but increased significantly at P6 in 2% FBS. Addition of VEGF did not cause any significant change in gene expression level. Conclusion: Adipose stromal cells had greater angiogenic potential when cultured in reduced serum conditions. VEGF did not enhance their angiogenic potential in 2% FBS-supplemented medium.

AB - Objectives: This study investigated effects of reduced serum condition and vascular endothelial growth factor (VEGF) on angiogenic potential of adipose stromal cells (ASCs) in vitro. Materials and methods: Adipose stromal cells were cultured in three different types of medium: (i) F12/DMEM (FD) supplemented with 10% FBS from passage 0 (P0) to P6; (ii) FD supplemented with 2% FBS at P6; and (iii) FD supplemented with 2% FBS plus 50 ng/ml of VEGF at P6. Morphological changes and growth rate of ASCs were recorded. Changes in stemness, angiogenic and endogenic genes' expressions were analysed using Real-Time PCR. Results: Adipose stromal cells changed from fibroblast-like shape when cultured in 10% FBS medium to polygonal when cultured in 2% FBS plus VEGF-supplemented medium. Their growth rate was lower in 2% FBS medium, but increased with addition of VEGF. Real-Time PCR showed that ASCs maintained most of their stemness and angiogenic genes' expression in 10% FBS at P1, P5 and P6, but this increased significantly in 2% FBS at P6. Endogenic genes expression such as PECAM-1, VE chaderin and VEGFR-2 decreased after serial passage in 10% FBS, but increased significantly at P6 in 2% FBS. Addition of VEGF did not cause any significant change in gene expression level. Conclusion: Adipose stromal cells had greater angiogenic potential when cultured in reduced serum conditions. VEGF did not enhance their angiogenic potential in 2% FBS-supplemented medium.

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