In vitro culture and differentiation of osteoblasts from human umbilical cord blood

Abstract

It is well accepted that human umbilical cord blood (UCB) is a source of mesenchymal stem cells (MSCs) which are able to differentiate into different cell phenotypes such as osteoblasts, chondrocytes, adipocytes, myocytes, cardiomyocytes and neurons. The aim of this study was to isolate MSCs from human UCB to determine their osteogenic potential by using different kinds of osteogenic medium. Eventually, only those MSCs cultured in osteogenic media enriched with vitamin D(2) and FGF9, were positive for osteocalcin by RT-PCR. All these cells were positive for alizarin red, alkaline phosphatase and Von Kossa. The results obtained from RT-PCR have confirmed that osteogenesis is complete by expression of the osteocalcin marker. In conclusion, vitamin D(2), at least in vitro, may replace vitamin D(3) as an osteogenic stimulator factor for MSC differentiation.

Culture and differentiation of osteoblasts on coral scaffold from human bone marrow mesenchymal stem cells

“…If we compare the data from the youngest donor, who is 18 years old, the density of his primary cells and the quantity of CFU-Fs is far more less than those from the oldest donor, who is 78 years old, with a ratio of 2.2/ml of cell density against 8.0/ml and a quantity of CFU-Fs of 34 against 110 (Table 1). A second point that comes out from this study is the equivalent function of vitamin D2 and vitamin D3 for osteoblast maturation in association with FGF9 (Jono et al 1998;Toai et al 2009). As already confirmed by several studies, vitamin D3 and FGF9 play an important …”

In vitro culture of Keratinocytes from human umbilical cord blood mesenchymal stem cells: the Saigonese culture

“…MSCs from human-UCB Human UCB is a significant source of hematopoietic stem cells and has been considered as a valid alternative for hematopoietic stem cell transplantation (Toai et al 2009;Lee et al 2004;Park et al 2006; Van de Ven et al 2007;Maurice et al 2007;Musina et al 2007;Sasaki et al 2008). MSCs from hUCB have been used in a wide range of diseases such as liver disorders, myocardial infarction, central nervous system condition or in degenerative pathologies such as diabetes, Crohn's disease, osteogenesis imperfect (OI), rheumatoid arthritis (RA) and osteoarthritis (OA) (Toai et al 2009;Lee et al 2004;Riordan et al 2007;Kogler and Wernet 2006;Kim et al 2004;Reddi 2007;Koblas et al 2005;De Bari and Dell'Accio 2007;Tuan and Chen 2006;Waese and Kandel 2007;Park et al 2006).…”

“…MSCs from hUCB have been used in a wide range of diseases such as liver disorders, myocardial infarction, central nervous system condition or in degenerative pathologies such as diabetes, Crohn's disease, osteogenesis imperfect (OI), rheumatoid arthritis (RA) and osteoarthritis (OA) (Toai et al 2009;Lee et al 2004;Riordan et al 2007;Kogler and Wernet 2006;Kim et al 2004;Reddi 2007;Koblas et al 2005;De Bari and Dell'Accio 2007;Tuan and Chen 2006;Waese and Kandel 2007;Park et al 2006). The most valuable potential of MSCs is their ability to switch into different cell phenotypes such as osteocytes, chondrocytes, adipocytes, hepatocytes, neurons, myocytes and keratinocytes with a great immunemodulatory and anti-inflammatory capacity that make them a tool for clinical applications (Toai et al 2009;Lee et al 2004;Goodwin et al 2001;Chamberlain et al 2007;Kim et al 2004;Bieback et al 2004;Musina et al 2007;Jang et al 2006;Rosada et al 2003; Van de Ven et al 2007;Maurice et al 2007;Tse and Laughlin 2005;Koc and Lazarus 2001;Chao et al 2004;Majhal et al 2006;Sasaki et al 2008;Stocum 2006).…”

“…The inconvenience of using bio-engineered materials for skin graft replacement is connected to the allogeneic origin of these cells hence these bio-products can only be used for wound coverage and not as a graft for tissue substitution (Markowicz et al 2005). Nevertheless, many studies have confirmed a conspicuous advantage of UCB engraftment related with a very low rate of transplant mortality and no increase of rejection or graft versus host disease (GVHD) due to a high rate of tolerance across 1 or 2 HLA-A, B and DR mismatches and a lower risk of infectious disease transmission (Van de Ven et al 2007;Toai et al 2009;Lee et al 2004;Riordan et al 2007;Kogler and Wernet 2006;Tse and Laughlin 2005). The idea to obtain keratinocytes from hUCB MSCs is mainly due to their particular ability to differentiate into different cell phenotypes and their immune-modulatory and anti-inflammatory nature that is crucial in the case of allograft procedures for skin regeneration ( Van de Ven et al 2007;Toai et al 2009;Lee et al 2004;Riordan et al 2007;Kogler and Wernet 2006;Tse and Laughlin 2005;Kamolz et al 2006).…”

“…Nevertheless, many studies have confirmed a conspicuous advantage of UCB engraftment related with a very low rate of transplant mortality and no increase of rejection or graft versus host disease (GVHD) due to a high rate of tolerance across 1 or 2 HLA-A, B and DR mismatches and a lower risk of infectious disease transmission (Van de Ven et al 2007;Toai et al 2009;Lee et al 2004;Riordan et al 2007;Kogler and Wernet 2006;Tse and Laughlin 2005). The idea to obtain keratinocytes from hUCB MSCs is mainly due to their particular ability to differentiate into different cell phenotypes and their immune-modulatory and anti-inflammatory nature that is crucial in the case of allograft procedures for skin regeneration ( Van de Ven et al 2007;Toai et al 2009;Lee et al 2004;Riordan et al 2007;Kogler and Wernet 2006;Tse and Laughlin 2005;Kamolz et al 2006). Human UCB have been shown to have a very limited number of graft lymphocytes and hUCB MSCs are able to secrete inhibitory cytokines such as IL10 and TFG-b whilst maintaining the ability of presenting antigens to T cells, a condition that eventually confirm a tolerogenic antigen capacity of this group of stem cells (Toai et al 2009;Lee et al 2004;Riordan et al 2007).…”