Human adipose-derived stem cells (HADSC) have demonstrated the capacity of differentiating into bone depending on the specific induction stimuli and growth factors. However, investigation on stem cell characteristic after osteogenic differentiation is still lacking. The goal of this study was to investigate the differential expression of sternness and osteogenic genes in non-induced HADSC compared with HADSC after osteogenic induction using quantitative Real Time RT-PCR. Our results showed that OCT-4, REX-1, FZD9, OSC, RUNX, and ALP were up regulated after osteogenic induction. This may indicated that HADSCs after osteogenic induction still possessed some stemness properties.

The objective of this study was to investigate the angiogenic potential of human chorion-derived stem cells (CDSC) cultured in medium containing bFGF and VEGF (EDM50). Total RNA was extracted from cells cultured in FD+10% FBS and EDM50. Quantitative RT-PCR was carried out to score the differential mRNA expression of genes involve in angiogenesis and endothelial differentiation. Our finding demonstrated that all angiogenic and endothelial associated genes were expressed higher in EDM50. Expression level of ANG-1, eNOS and VEGFR2 were significantly higher in EDM50 compared to FD+10% FBS. Our results suggested that human CDSC cultured in EDM50 can be used for angiogenesis purpose in regenerative medicine.

A major factor limiting survival following extensive thermal injury is insufficient availability of donor sites to provide enough skin for the required grafting procedures. Limitation of autologous grafting promotes the usage of allograft skin substitutes to promote wound healing. Here, we investigated the wound healing potential of allograft single layered tissue engineered skin which comprises of either keratinocytes (SLTES-K) or fibroblast (SLTES-F) with fibrin as the delivery system. Results from gross and microscopic evaluation showed our single layered tissue engineered skin constructed with keratinocytes or fibroblast after gamma radiation with the dosage of 2Gy could serve as allograft for the treatment of skin loss.

The aim of the study is to determine the neuronal and glial gene expression of cultured human amniotic epithelial cells (HAECs) in serial passages. HAECs obtained from human term placentae were cultured in F12:DMEM (1:1) + 10% FBS +10ng/ml EGF in serial passages. Quantitative RT-PCR was used to assess the gene expression analysis. The results showed that the cultured HAECs expressed the neural stem cell genes (Nestin, NSE and Vimentin), mature neuronal genes (TH, MAP-2, beta-III-tubulin and NFM) and glial genes (CNPase, MBP and Olig). These neural stem cell genes increased with serial passages while the genes expression for mature neuronal and glial cells were downregulated. These results suggested that HAECs may promote or involve in neurogenesis and gliagenesis.

The aim of the study is to evaluate the stemness gene expression of cultured human amniotic epithelial cells (HAECs) in serial passages. HAECs obtained from human term placentae were cultured in F12:DMEM(1:1) + 10% FBS +10ng/ml EGF in serial passages (P0, P1, P2 and P4). Quantitative RT-PCR was used to assess the gene expression analysis. The results showed that cultured HAECs expressed and downregulated the stemness genes expression for Oct-4, Sox-2, Nanog3, FGF4, Rex-1, FZD-9, BST-1 ABCG2. However, vimentin and nestin gene expression were upregulated. The results suggested that cultured HAECs may have pluripotent and multipotent properties.

Angiogenic induction was made to promote angiogenesis by differentiating stem cells towards endothelial cells. However, the stemness property of induced cells has not been revealed yet. Hence, we aim to evaluate the differential mRNA expression of stemness genes in human chorion-derived stem cells (CDSC) after being cultured in EDM50 comprised bFGF and VEGF. Results indicated that CDSC cultured in EMD50 expressed significantly higher mRNA level of Sox-2, FZD9, BST-1 and Nestin. In addition Oct-4, FGF-4 and ABCG-2 were also upregulated. Our finding suggested that CDSC after angiogenic induction enhanced its stem cell properties. This could be contributed for the mechanism of stem cell therapy in ischemic problem.

It is crucial to know whether stem cells retain its stemnness properties after advance in vitro manipulation. The objective of this study was to investigate the stemness gene expression of human adipose tissue derived stem cells (ADSCs) in long-term culture using quantitative RT-PCR technique. Our data showed that the expression level of Sox-2, Rex-1, FGF-4, Nanog, Nestin, BST-1, FZD-9 and Oct-4 were decreased gradually in long-term culture. This could mean that the ability of ADSCs to differentiate into other cell lineages reduce after extensive culture.

Stress-induced premature senescence (SIPS) model is in vitro model of cellular aging. In this study, apoptosis was evaluated in SIPS model and in replicative senescent fibroblasts. We also compared the activity of senescence-associated beta-galactosidase (SA-beta gal) as a biomarker of cellular aging. Our results suggested that SIPS model and senescent fibroblasts might share similar mechanism of aging and apoptosis pathway.

This study was conducted to explore the feasibility of culturing conjunctiva epithelial cells in serum-free and feeder layer-free culture system with regard to the cell morphology and immunocytochemistry of the rabbit bulbar, fornix and palpebral conjunctiva epithelia. The results showed that epithelium cells from all the three conjunctiva regions can be cultured in a serum-free and feeder layer-free environment. We obtained highest epithelial growth from fornix region with minimum invasion of fibroblast cells compared to other area. All cultured cells were stained positive for cytokeratin 19 and MUC5AC and negative for cytokeratin 3. These findings suggested that fornix was a better source of cells for the development of tissue engineered conjunctiva for future clinical application.

A potential cure for hearing loss would be to regenerate hair cells by stimulating cells of the damaged inner ear sensory epithelia to proliferate and differentiate into hair cells. Here, we investigated the possibility to isolate, culture-expand and characterize the cells from the cochlea membrane of adult mice. Our results showed that the cultured cells isolated from mouse cochlea membrane were heterogenous in nature. Morphologically there were epithelial like cells, hair cell like, nerve cell like and fibroblastic cells observed in the culture. The cultured cells were immunopositive for specific hair cell markers including Myosin 7a, Calretinin and Espin.