3-D spheroid culture of bone marrow mesenchymal stem cell of rhesus monkey with improved multi-differentiation potential to epithelial progenitors and neuron in vitro
Article first published online: 23 MAY 2011
© 2011 The Authors. Clinical and Experimental Ophthalmology © 2011 Royal Australian and New Zealand College of Ophthalmologists
Clinical & Experimental Ophthalmology
Volume 39, Issue 8, pages 808–819, November 2011
How to Cite
Jing, Y. and Jian-xiong, Y. (2011), 3-D spheroid culture of bone marrow mesenchymal stem cell of rhesus monkey with improved multi-differentiation potential to epithelial progenitors and neuron in vitro. Clinical & Experimental Ophthalmology, 39: 808–819. doi: 10.1111/j.1442-9071.2011.02560.x
- Issue published online: 3 NOV 2011
- Article first published online: 23 MAY 2011
- Accepted manuscript online: 15 MAR 2011 05:16AM EST
- Received 14 November 2010; accepted 11 February 2011.
- 3-D spheroid culture system;
- epithelial progenitor;
- mesenchymal stem cell;
- rhesus monkey
Background: Bone mesenchymal stem cells are an attractive source of cells with potential applications in ocular regenerative medicine. However, the low differentiation efficiency in the traditional two-dimensional (2-D) culture system limits their application for clinical therapy. Here, we describe a simple and innovative 3-D culture environment and assess the potential for bone mesenchymal stem cells to differentiate into a variety of cell types in the 3-D system.
Methods: Bone mesenchymal stem cells of rhesus monkey were isolated and cultured using a density gradient centrifugation and adherence screening method. Cells at passage three were cultured by hanging drop and formed spheres. After 3 days, the spheres were collected and plated onto culture plates and maintained in a floating state by a rotary method for 10 days. Under appropriate induction conditions, the sphere cells were induced into adipocytes, osteoblasts, epithelial progenitors and neuronal cells. Differentiated cells were identified by histochemical staining, immunofluorescence and reverse transcription-polymerase chain reaction.
Results: Bone mesenchymal stem cells of rhesus monkey in the 3-D spheroid culture system acquired improved efficient multipotency for not only adipogenic and osteogeneic differentiation, but ectodermal epithelial progenitor-like cells and neuron-like cells compared with the 2-D culture system.
Conclusion: Our 3-D spheroid culture system provides a useful technique to gain insight into the mechanisms of bone mesenchymal stem cells differentiation, and may have potential applications for the treatment of ocular surface and optic nerve disease.