The Bone Marrow Mesenchymal Cells are important for the growth and differentiation of progenitor of haematopoietic cells.
The immortalized Human Bone Marrow Mesenchymal Cells was created by infecting human primary mesenchymal cells with retrovirus containing hTERT and GFP genes separated by IRES under the control of MSCV LTR (pCIneo-hEST2-HA).
The Immortalized Bone Marrow Mesenchymal cells can support growth of leukaemic cell lines 380, REH, OP-1 and 697, in addition to support the expansion of CD34+ cells.
This cell line is useful not only in supporting the growth of haematopoietic cells, but can also be used in studies concerning bone marrow microenvironment and environmental cues that regulate haematopoiesis.
Immortalization Method
Serial passaging and transduction with recombinant VSV-G retrovirus carrying hTERT and GFP genes
BioSafety Level
BSL-II
Species
Human
Source Organ
Bone Marrow
Growth Properties
Adherent
Morphology
Spindle shaped
Population Doubling
35-45 hours
Cell Seeding Density
10,000 - 20,000 cells/cm²; Recommended split ratio is 1:2 to 1:3
Markers
CD44 and CD73.
These cells also express markers of osteoblasts (alkaline phosphatise) and chondrocytes (incorporation of toluidine blue dye)
Propagation
The base medium for this cell line is Prigrow II medium (Cat. No. ABM-TM002). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ABM-TM999)* to a final concentration of 10%, hydrocortisone to 10-6 mol/L and Penicillin/Streptomycin Solution (ABM-G255) to a final concentration of 1%.
Quality Control
1. Immunofluorescence and RT-PCR were used to confirm the presence hTERT expression.
2. PCR based TRAP was used to assay telomerase activity.
3. Neo concentration not determined for selection use but cells were sorted for GFP+ populations.
Shipping Condition
Dry Ice
Storage Condition
liquid nitrogen or -180°C