|
Print Version |
| BioSafety Level |
II |
| Organism |
Homo sapiens |
| Source Organ |
Bone marrow |
| Growth Properties |
Adherent |
| Morphology |
Spindle-shaped |
| Population Doubling |
Doubling time of approximately 100 hours |
| Recommended Seeding Density |
Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions. |
| Markers |
CD44, NT5E, THY1, ENG, CD90 |
| Applications |
For Research Use Only |
| Immortalization Method |
Serial passaging and transduction with recombinant lentiviruses carrying hTERT gene (pBabe-puro-hTERT) |
| Description |
Human mesenchymal stromal cells have extensive potential in research: uses for these including tissue repair and tissue engineering, gene therapy, the study of mesenchymal tumor oncogenes, and the analysis of cell fate determination and differentiation. The iMSC#3 cells express markers found in both primary human mesenchymal stromal cells and the multipotent form: CD90, CD44, NT5E, THY1, ENG, alanyl aminopeptidase (ANPEP), integrin beta I (ITGB1), integrin alpha 5 (ITGA5), major histocompatibililty complex, class I, A and B (HLA-A and B), and activated leukocyte cell adhesion molecule (ALCAM). These cells have also shown the ability to differentiate into adipocytes and osteoblasts. Furthermore, the cells have been verified for normal copy number, and lacks chromosomal aberrations and tumorigenicity. |
| 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 III medium available from ABM (TM003). To make the completed growth medium, add the following components to the base medium: 10% heat-inactivated fetal bovine serum, and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. |
| Preservation |
1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.
2. Storage Temperature: Liquid nitrogen vapour phase. |
| Quality Control |
(1) Human telomerase reverse transcriptase expression was verified with quantitative PCR; (2) Surface markers were determined with flow cytometry; (3) Gene expression profile was analysed using the Illumina WG-6 v2 Expression BeadChip and Illumina HumanHT12 v4; (4) Adiopocyte differentiation was detected with Oil Red O staining, while osteoblast differentiation was detected by staining cells with Alizarin Red and ALPL and quantification; (5) DNA methylation profiling was performed using the Illumina HumanMethylation450 BeadChip. Copy number verification was performed with high-resolution mapping using Affymetrix Genome-Wide Human SNP Array 6.0.; (6) Karyotype analysis was performed by arresting cells during metaphase and subjecting chromosomes to G-banding and subsequent imaging; (7) Tumorgenicity was verified in vivo with subcutaneous injection into immunodeficient athymic mice; (8) miRNA expression profile was determined using high-throughput sequencing. |
| 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. |
