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Immortalized-Human-Bone-Marrow-Mesenchymal-Cells---hTERT

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Immortalized-Human-Bone-Marrow-Mesenchymal-Cells---hTERT

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Immortalized-Human-Bone-Marrow-Mesenchymal-Cells---hTERT

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Bone Marrow
Growth Properties	Adherent
Morphology	spindle-shaped
Population Doubling	39 hours
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	CD44 and CD73. These cells also express markers of osteoblasts (alkaline phosphatise) and chondrocytes (incorporation of toluidine blue dye)
Applications	For research only. The Immortalized Human Bone Marrow Mesenchymal Cells- hTERT is unique that 1. The cells retained inhibition of proliferation by cell-cell contact and did no form foci in soft agar; 2. The cells exhibit no karyotypic abnormalities; 3. The cells retained identical phenotype to that of their primary counterparts; 4. The cells can be induced to differenciate into osteoblasts and chondrocytes.
Immortalization Method	Serial passaging and transduction with recombinant VSV-G retrovirus carrying hTERT and GFP genes
Description	The Bone Marrow Mesenchymal Cells are important for the growth and differentiation of primitive haematopoietic cells, as well as for maintenance of leukaemic cells. The immortalized cell was created by infecting primary mesenchymal cells with retrovirus containing hTERT and GFP genes separated by IRES under the control of MSCV LTR. 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.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow II medium available in ABM, Cat. No. TM002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10%, hydrocortisone to 10^-6 mol/L and Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO₂), 5%.
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.
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.