Immortalized Human Cord Blood Mesenchymal Stromal Cells with RFP (cbMSC-hTERT-RFP)

Owing to its multiple differentiation abilities, the Mesenchymal stromal cells (MSCs) holds the promise in tissue engineering and regenerative medicine, and displays important clinical potential for cell therapy. The Immortalized Human Cord Blood Mesenchymal Stromal Cells with RFP (cbMSC-hTERT-RFP) retains MSC specific surface markers (CD29, CD73, CD105, CD44 and HLA-ABC) as well as its differentiation capacities into cells of the mesodermal lineage, such as osteoblasts and adipocytes. Karyotype analysis and CGH analysis show no chromosomal abnormality nor genomic deletion or amplifications. In addition, transplantation of cbMSC-hTERT-RFP into the rat model with TBI demonstrates that these cells are readily attracted to the injury area and thus provides a valuable cell-based tool forin vivostem cell research and translational study.

Immortalization Method: Serial passaging and transfection with Human Telomerase Reverse Transcriptase (hTERT) expression vector pGRN145

BioSafety Level: II

Species: Human

Source Organ: Umbilical cord blood

Growth Properties: Adherent

Morphology: Fibroblast-like

Population Doubling: Population doubling time of 23.8 ± 1.9 h

Seeding Density: Subculture ratio of 1:3 to 1:5

Markers: CD90, CD34, CD44, CD45, CD166, CD117,CD73, CD105, CK18 and CK19

Donor Age: N/A

Donor Gender: Female

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow III medium available at abm (TM003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 20%, 4 ng/ml bFGF (Cat. Z101455), and Penicillin/streptomycin at a final concentration of 1%. It is recommended to renew medium every 2 to 3 days.
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) RT-PCR was used to confirm the expression of the hTERT transgene in the immortalized cells; 2) TRAP assay was performed to determine the telomerase activity; 3) Flow cytometry was used to detect surface markers such as CD29, CD73, CD105, CD44 and HLA-ABC; 4) G-banding was used for karyotype analysis; 5) Tumuorigenicity assay was performed to evaluate the cells’ tumorigenic potential.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C