Bone marrow mesenchymal stem cells differentiate into urothelial cells and the implications for reconstructing urinary bladder mucosa

Abstract

To determine the ability of cultured bone marrow-derived mesenchymal stem cells (BMSCs) to differentiate into functional urothelium. BMSCs were isolated from the long bones of aborted fetal limbs by Percoll density gradient centrifugation and characterized by flow cytometry. Human fetal urinary bladders were cut into small pieces and cultured for 3-5 days until the growth of urothelial cells was established. BMSCs were then cocultured with neonatal urothelial cells and subsequently evaluated for antigen expression and ultramicrostructure, by immunocytochemistry and electron microscopy, respectively. A subset of BMSCs expressed the differentiation marker CD71. The BMSC markers CD34, CD45, and HLA-DR were barely detectable, confirming that these cells were not derived from hematopoietic stem cells or differentiated cells. In contrast, the stem cell markers CD29, CD44, CD105, and CD90 were highly expressed. BMSCs possessed the ability to differentiate into a variety of cellular subtypes, including osteocytes, adipocytes, and chondrocytes. The shapes of BMSCs changed, and the size of the cells increased, following in vitro coculture with urothelial cells. After 2 weeks of coculture, immunostaining of the newly differentiated BMSCs positively displayed the urothelialspecific keratin marker. Electron microscopy revealed that the cocultured BMSCs had microstructural features characteristic of epithelial cells. Pluripotent BMSCs can transdifferentiate into urothelial cells in response to an environment conditioned by neonatal urothelial cells, providing a means for the time-, labor-and cost-effective reconstruction of urinary bladder mucosa.

A tissue-engineered conduit for urinary diversion using bone marrow mesenchymal stem cells and bladder acellular matrix

“…Previous reports have revealed that BMSCs can transform into urothelium-like cells following either treatment with conditioned medium or by co-culturing them with urothelial cells. BMSCs have been shown to express urothelium-specific genes and proteins following these treatments [12,18]. Our findings also demonstrated the feasibility of obtaining urothelium-like cells from BMSCs.…”

A fibrin/hyaluronic acid hydrogel for the delivery of mesenchymal stem cells and potential for articular cartilage repair

“…CD44 has also been documented as a phenotypical MSC marker [61-63] and was also assayed for. CD105-PE, CD73-PerCP, CD90-APC, CD44-FITC and CD34-FITC surface markers were assessed using an Accuri C6 Flow Cytometer.…”

The Current Use of Stem Cells in Bladder Tissue Regeneration and Bioengineering

“…Histological evaluation revealed bladder tissue formation, and immunostaining verified uroplakin expression, confirming urothelial differentiation [48]. This was similarly shown when human bone marrow-derived mesenchymal stem cells were co-cultured with human urothelial cells and developed urothelial cell features [21]. Tian et al further revealed that MSCs can differentiate into urothelium when only cultured in conditioned medium derived from bladder cell culture optimized for urothelial differentiation, suggesting the feasibility of cell free differentiation systems [22].…”