Immortalized Human Bone Marrow Mesenchymal Stromal Cells - hTERT (iMSC3) | T0529

Immortalized Human Bone Marrow Mesenchymal Stromal Cells - hTERT (iMSC3) | T0529

Immortalization Method:

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

BioSafety Level:

II

Organism:

Homo sapiens

Species:

Human

Source Organ:

Bone marrow

Organ Type:

Blood

Growth Properties:

Adherent

Morphology:

Spindle shaped

Passage Number:

N/A

Population Doubling:

45 - 50 hours

Seeding Density:

20,000 cells/cm10²

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 . 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 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 -180C