Murine mesenchymal stem cells isolated by low density primary culture system

Abstract

Murine mesenchymal stem cells (mMSC) and the difficult task of isolation and purification of them have been the subject of rather extensive investigation. The present study sought to isolate these cells from two different mouse strains, one outbred and the other inbred, primarily through a relatively simple but novel approach, the most important feature of which was the low density primary culture of bone marrow cells. For this purpose, mononuclear cells from either NMRI or BALB/c bone marrow were plated at about 500 cells per well of 24-well plates and incubated for 7 days. At this point, the fibroblastic clones that had emerged were pooled together and expanded through several subcultures. To investigate the mesenchymal nature, we differentiated the cells into the osteoblastic, chondrocytic and adipocytic lineages in different subcultures up to passage 10. According to the results, 1 week after culture initiation, several clones each comprising several fibroblastic cells appeared in each plate. The cells from different passages were capable of differentiating into corresponding skeletal tissues. In the present investigation, the best culture condition for maximum proliferation and also the expression of certain surface marker on isolated cells were examined. In this term the two murine strains showed some differences.

The isolation and characterization of putative mesenchymal stem cells from the spiny mouse

“…Similarly, isolated MSCs can be directed to differentiate in vitro, by culture with lineage specific differentiation medium, to differentiate down the osteogenic, adipogenic and chondrogenic lineages (Gregory et al 2005). The fibroblast-like cells isolated from the spiny mice in this study exhibited multilineage differentiation potential with the capacity for adipogenic, chondrogenic and osteogenic differentiation, as demonstrated for MSCs isolated from other species (Bosnakovski et al 2006;Horwitz et al 2005;Izadpanah et al 2005;Eslaminejad et al 2006;Tuli et al 2003).…”

“…They also exhibited characteristic morphological features of MSCs, such as plastic adherence and spindle-shaped fibroblastic morphology (Eslaminejad et al 2006;Izadpanah et al 2005;Lange et al 2005;Tang et al 2006). MSCs differentiate by default in vivo into cell lineages of mesenchymal tissues, including bone, cartilage, fat, muscle, and myelosupportive stroma (Kyriakou et al 2006).…”

Induced in vitro differentiation of neural-like cells from human exfoliated deciduous teeth-derived stem cells

“…Osteogenic and adipogenic differentiation of DPSCs were induced as previously described (Eslaminejad et al, 2006). To achieve osteogenic differentiation, cells were seeded at a density of 310 3 cells/cm 2 and then cultured for ten days in the presence of 10 nM dexamethasone, 50 g/ml ascorbic acid 2-phosphate and 10 mM -glycerophosphate.…”

“…Adipogenesis was assayed by staining of intracellular lipid droplets with oil red O (Sigma) as previously described (Eslaminejad et al, 2006).…”

Characterization of fibroblast-like cells from the rat olfactory bulb

“…The cultured cells were incubated in osteogenic conditioned medium (Eslaminejad et al, 2006;Nadri et al, 2007). Briefly, DMEM was supplemented with 10 mM β-glycerol phosphate (Sigma), 50 µg/ml ascorbate-2-phosphate (Sigma) and 10 -7 M dexamethasone (Sigma) (Nadri et al, 2007).…”