Adipose-Derived Stem Cell Coculturing Stimulates Integrin-Mediated Extracellular Matrix Adhesion of Melanocytes by Upregulating Growth Factors

Coculture with adipose-derived stem cells (ADSCs) can stimulate proliferation and migration of melanocytes. To enhance outcomes of skin disorders caused by melanocyte loss or death, mixed transplantation with ADSCs has been suggested. However, role of cocultured ADSCs in proliferation and migration of melanocytes remains unclear. This study determined the effect of ADSCs on production of growth factors and expression levels of intergrins in primary culture of adult human melanocytes with or without ADSCs and in nude mice grafted with such melanocytes. Higher amounts of growth factors for melanocytes, such as bFGF and SCF were produced and released from ADSCs by coculturing with melanocytes. Relative levels of integrins β1, α5, and α6 as well as adhesion to fibronectin and laminin were increased in melanocytes cocultured with ADSCs. Such increases were inhibited by neutralization of bFGF or SCF. Relative levels of bFGF, SCF and integrins were increased in nude mice skin after grafting with melanocyte+ADSC cocultures. Collectively, these results indicate that ADSCs can stimulate proliferation and migration of melanocytes by increasing expression of integrins in melanocytes through upregulation of production/release of melanocyte growth factors such as bFGF and SCF.

The effects of electromagnetic fields on osteogenesis during co-culture of bone marrow mesenchymal stem cells with osteoblasts / 中华物理医学与康复杂志

Objective To explore the effects of electromagnetic fields (EMFs) on osteogenesis during co-culture of bone mesenchymal stem cells (BMSCs) with osteoblasts in rats.Methods BMSCs and osteoblasts were isolated from Sprague-Dawley rats and cultured.Sub-cultured osteoblasts and BMSCs were seeded in transwell cell-culture-chamber polyester inserts to establish the co-culture system.The co-cultures were then randomly divided into a normal co-culture group and a group exposed to an EMF.Single-cultured BMSCs and osteoblasts were set as a single culture group.The EMF group was exposed to an EMF for 4 hours per day.On the 14th day,cell culture plates or inserts were randomly selected for total RNA extraction and measurement of the mRNA expression levels of Runt-related transcription factor 2,transcription factor 7,alkaline phosphatase,collagen type Ⅰ,bone morphogenetic protein 2 (BMP-2) and bone gamma-carboxyglutamate protein (Osteocalcin gene,OC gene) using real-time PCR assays.Cell culture dishes or inserts were also randomly chosen for Alizarin red staining to detect mineralized nodules.Results The level of osteogenic gene expression in single-cultured BMSCs and osteoblasts was low,while it was much higher in the co-culture group.The level of gene expression in the EMF-exposed and co-cultured group was even higher.Alizarin red staining also showed that calcium mineralized modules had increased in the stimulated,co-cultured system compared with the unstimulated,co-cultured cells.Conclusion EMF exposure can promote osteogenic differentiation of BMSCs and osteoblasts when they are co-cultured.BMP2-mediated cellular interaction might play an important role in osteogenic differentiation induced by EMF exposure.

Successful pregnancies using frozen thawed testicular sperm in ICSI program / 대한산부인과학회잡지

There are not much reports concerning with clinical results using frozen-thawed testivular sperm in ICSI program. It is speculated that the necessity of cryopreservation of testicular sperm to avoid repeating surgical procedure for obtaining sperm for ICSI. This study was carried out to confirm whether frozen-thawed testicular sperm could be fertilized and pregnancy could be achieved using embryos fertilized with frozen-thawed testicular sperm in ICSI program or not. Testicular sperm obtained from obstructive- or non-obstructive azoospermia patients were co-cultured for 3 days with Vero cells to improve sperm motility. By co-culturing with Vero cells for 3 days, O-ll% of sperm motility after thawing increased up to 8-42% after co-culturing. ICSI was performed using frozen-thawed, and co-cultured sperm with 66 oocytes obtained from 8 patients and 62 oocytes were survived and 49(79.0%) oocytes were fertilized normally. Embryo transfer was possible in 7 out of 8 patients, and pregnancy was achieved in 6 patients(85.7%). These results indicated that not only fresh testicular sperm but frozen-thawed testicular sperm can be used in ICSI program.