Exp Mol Med.  2012 Apr;44(4):303-309. 10.3858/emm.2012.44.4.034.

Vascular differentiation of multipotent spermatogonial stem cells derived from neonatal mouse testis

• Im JE
• Song SH
• Kim JY
• Kim KL
• Baek SH
• Lee DR
• Suh W

• KMID: 1365907
• DOI: http://doi.org/10.3858/emm.2012.44.4.034

Abstract

We previously reported the successful establishment of embryonic stem cell (ESC)-like multipotent spermatogonial stem cells (mSSCs) from neonatal mouse testis. Here, we examined the ability of mSSCs to differentiate into vascular endothelial cells and smooth muscle cells, and compared to that of mouse ESCs. We used real-time reverse transcriptase polymerase chain reaction and immunohistochemistry to examine gene expression profiles of mSSCs and ESCs during in vitro vascular differentiation. Both mSSCs and ESCs exhibited substantial increase in the expression of mesodermal markers, such as Brachyury, Flk1, Mesp1, Nkx2.5, and Islet1, and a decrease in the expression of pluripotency markers, such as Oct3/4 and Nanog during the early stage of differentiation. The mRNA levels of vascular endothelial (VE)-cadherin and CD31 gradually increased in both differentiated mSSCs and ESCs. VE-cadherin- or CD31-positive cells formed sprouting branch-like structures, as observed during embryonic vascular development. At the same time, vascular smooth muscle cell-specific markers, such as myocardin and alpha-smooth muscle actin (SMA), were also highly expressed in differentiated mSSCs and ESCs. Immunocytochemical analysis revealed that the differentiated cells expressed both alpha-SMA and SM22-alpha proteins, and exhibited the intracellular fibril structure typical of smooth muscle cells. Overall, our findings showed that mSSCs have similar vascular differentiation abilities to those of ESCs, suggesting that mSSCs may be an alternative source of autologous pluripotent stem cells for vascular regeneration.

Keyword

cell differentiation; endothelial cells; multipotent stem cells; myocytes, smooth muscle; spermatogonia

MeSH Terms

Animals
Animals, Newborn
Biological Markers/metabolism
Cell Differentiation/physiology
Embryonic Stem Cells/cytology/physiology
Endothelial Cells/*cytology/physiology
Gene Expression
Gene Expression Profiling
Humans
Immunohistochemistry
Male
Mice
Muscle, Smooth, Vascular/*cytology/physiology
Myocytes, Smooth Muscle/*cytology/physiology
Pluripotent Stem Cells/*cytology/physiology
Real-Time Polymerase Chain Reaction
Spermatogonia/*cytology/physiology
Testis/*cytology/physiology