Korean J Urol.  2011 Dec;52(12):852-857.

Comparison of Human Muscle-Derived Stem Cells and Human Adipose-Derived Stem Cells in Neurogenic Trans-Differentiation

• Kwon EB
• Lee JY
• Piao S
• Kim IG
• Ra JC
• Lee JY

Abstract

PURPOSE
Erectile dysfunction (ED) remains a major complication from cavernous nerve injury during radical prostatectomy. Recently, stem cell treatment for ED has been widely reported. This study was conducted to investigate the availability, differentiation into functional cells, and potential of human muscle-derived stem cells (hMDSCs) and human adipose-derived stem cells (hADSCs) for ED treatment.
MATERIALS AND METHODS
We compared the neural differentiation of hMDSCs and hADSCs. Human muscle and adipose tissues were digested with collagenase, followed by filtering and centrifugation. For neural induction, isolated hMDSCs and hADSCs were incubated in neurobasal media containing forskolin, laminin, basic-fibroblast growth factor, and epidermal growth factor for 5 days. Following neural induction, hMDSCs and hADSCs were differentiated into neural cells, including neurons and glia, in vitro.
RESULTS
In neural differentiated hMDSCs (d-hMDSCs) and differentiated hADSCs (d-hADSCs), neural stem cell marker (nestin) showed a significant decrease by immunocytochemistry, and neuronal marker (beta-tubulin III) and glial marker (GFAP) showed a significant increase, compared with primary hMDSCs and hADSCs. Real-time chain reaction analysis and Western blotting demonstrated significantly elevated levels of mRNA and protein of beta-tubulin III and GFAP in d-hADSCs compared with d-hMDSCs.
CONCLUSIONS
We demonstrated that hMDSCs and hADSCs can be induced to undergo phenotypic and molecular changes consistent with neurons. The neural differentiation capacity of hADSCs was better than that of hMDSCs.

Keyword

Adipose tissue; Cell differentiation; Muscles; Neurons; Stem cells

MeSH Terms

Adipose Tissue
Blotting, Western
Caves
Cell Differentiation
Centrifugation
Collagenases
Epidermal Growth Factor
Erectile Dysfunction
Forskolin
Humans
Immunohistochemistry
Laminin
Male
Muscles
Neural Stem Cells
Neuroglia
Neurons
Prostatectomy
RNA, Messenger
Stem Cells
Tubulin
Collagenases
Epidermal Growth Factor
Forskolin
Laminin
RNA, Messenger
Tubulin

Figure

• 

  FIG. 1 Morphology and immunocytochemistry of nestin, β-tubulin III, and GFAP before and after human muscle-derived stem cells (hMDSCs) and human-adipose tissue derived stem cells (hADSCs) differentiation. Morphology and double fluorescent immunocytochemistry for nestin (green), β-tubulin III (green), and GFAP (green) shown in (A) hMDSCs, differentiated hMDSCs (d-hMDSCs), and (B) hADSCs, differentiated hADSCs (d-hADSCs). Nuclei are stained with DAPI (blue). Magnification is ×100.

• 

  FIG. 2 mRNA expression of nestin, β-tubulin III, and GFAP before and after human muscle-derived stem cells (hMDSCs) and human-adipose tissue derived stem cells (hADSCs) differentiation. (A) Nestin, (B) β-tubulin III, and (C) GFAP mRNA levels in hMDSCs, differentiated hMDSCs (dhMDSCs), hADSCs, and differentiated hADSCs (d-hADSCs) were quantified by real-time chain reaction analysis analysis. GAPDH was used as the internal control gene. Data are the mean±SD of three different cultures of stem cells (ap<0.01).

• 

  FIG. 3 Protein expression of nestin, β-tubulin III, and GFAP before and after human muscle-derived stem cells (hMDSCs) and human-adipose tissue derived stem cells (hADSCs) differentiation. (A) Nestin, β-tubulin III, and GFAP protein expression in hMDSCs, differentiated hMDSCs (d-hMDSCs), hADSCs, and differentiated hADSCs (d-hADSCs) were determined by Western blot analysis. Blots were probed with β-actin to show equal loading. The bar graphs show the relative levels of (B) nestin, (C) β-tubulin, and (D) GFAP protein in hMDSCs, d-hMDSCs, hADSCs, and d-hADSCs normalized to β-actin (n=3). Mean±SD, ap<0.01.

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