Immortalized Human Bone Marrow Mesenchymal Stromal Cells - hTERT (iMSC3)

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.

Immortalization Method: Serial passaging and transduction with recombinant retroviruses carrying hTERT gene (pBabe-puro-hTERT)

BioSafety Level: II

Species: Human

Source Organ: Bone marrow

Growth Properties: Adherent

Morphology: Spindle shaped

Population Doubling: 45 - 50 hours

Seeding Density: 20,000 cells/cm²

Markers: CD44, NT5E, THY1, ENG, CD90

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: 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: heat-inactivated fetal bovine serum to a final concentration of 10%, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.

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.(9) Puromycin selection used (2 μg/mL) however not needed for long term culture.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C