Identification of cultured progenitor cells from human marrow stroma

Abstract

The marrow stromal cells (MSC) are essential for regulation of bone remodeling and hematopoiesis. It is of prime importance to isolate MSC and to expand the proliferating cells ex vivo. In this study, we analyzed cultured MSC for various cellular parameters, including cell morphology, cell cycle, and expression of cell surface antigens by flow cytometry. MSC were divided based on cell size to small (S-cells) and large (L-cells) and were visualized by light and electron microscope. The S-cells were proliferating cells correlated with G0/G1 phase of cell cycle, and expressed cFOS. The expression of surface markers CD-34, -44, -51, -61, -62E, -62P, -62L was quantified using flow cytometry. CD-44 was ubiquitously expressed by S and L cells, CD-51 and -61 were expressed by 30%-38% of S-cells. CD-34 and -62 expressed 20% positive of the analyzed cells that were of the proliferating progenitors (S-cells). This study enables the identification of subpopulations from MSC with special attention paid to the proliferating cells from ex vivo cultures of marrow stroma.

Neither Notch1 Expression nor Cellular Size Correlate with Mesenchymal Stem Cell Properties of Adult Articular Chondrocytes

“…Recently, it has been shown that articular cartilage in rabbits grows appositionally until puberty implying a progenitor cell population located at the cartilage surface in young animals [Hunziker et al, 2007]. This zone consists of small cells and it has been demonstrated in other tissues that small cells express known markers for progenitor cells [Shur et al, 2002]. Additionally, surface cells are characterized by their Notch1 expression [Dowthwaite et al, 2004].…”

“…One characteristic of the cells in this zone is their small size compared to cells found in deeper layers of articular cartilage [Hidaka et al, 2006]. Small cellular size is characteristic for mesenchymal progenitor cells, while larger cells represent a more differentiated state [Shur et al, 2002]. The small cells located in the surface zone of articular cartilage might thus have progenitor cell characteristics with improved capacity to produce hyaline cartilage.…”

Increased signaling through p62 in the marrow microenvironment increases myeloma cell growth and osteoclast formation

“…by guest www.bloodjournal.org From Because marrow stromal cells are a heterogenous cell population and comprise fibroblast-like cells, macrophages, and endothelial cells, we next determined if the differences between MM patient-derived and normal stromal cells were due to a change in the distribution of cell types present in the stromal cell preparations, such as enrichment or loss of a particular cell type that affected the capacity of the stromal cell preparations to stimulate the growth of MM cells. Both MM and normal stromal cells contained similar percentages of cells that expressed CD63 (88.5 Ϯ 1.4 vs 88.4 Ϯ 1.8, mean Ϯ standard deviation [SD] for healthy vs MM patients, n ϭ 5) and CD51 (86 Ϯ 0.5 vs 90 Ϯ 3, mean Ϯ SD for healthy vs MM patients, n ϭ 5) 23,24 and that had low expression of CD14 (11 Ϯ 1 vs 12 Ϯ 0.3, mean Ϯ SD for healthy vs MM patients, n ϭ 5; Figure 1D). The hematopoietic cell marker for the monocyte lineage (CD11b) was not detected in either stromal cell preparation (data not shown).…”

MS‐KIF18A, a kinesin, is associated with estrogen receptor

“…Ex vivo primary cultured human mesenchymal marrow cells (MSC) [Shur et al, 2002] and MBA-15, a mouse marrow stromal derived cell line [Benayahu et al, 1989] were cultured in growth medium containing Dulbecco's modified essential medium (DMEM) (Gibco) with the addition of 10% heat-inactivated fetal calf serum (FCS, Biological Industries, Bet-Haemek, Israel). COS-7 cells were grown in DMEM medium supplemented with 5% FCS.…”