OP9

€375.00*

Product number: 305174

Description	OP9, a macrophage-derived embryonic stem cell line, was isolated from the stroma of a mouse embryo. With their fibroblast-like morphology and origin from the bone marrow stromal cells, OP9 cells have proven to be instrumental in advancing our understanding of cell therapy, regenerative medicine, and immunomodulation. It shares an identical immunophenotype with standard mouse mesenchymal stem cells (MSCs). The cell line was cultured from the newborn calvaria of osteopetrosis mutant mice, which lack functional macrophage colony-stimulating factor (M-CSF). Specific markers such as CD140a+, CD140b+, α-SMA+, and Calponin+ suggest a perivascular origin for these cells. Morphologically, OP9 cells exhibit a distinct bipolar, fibroblast-like appearance. Their characteristics have been extensively studied using immunofluorescence staining and flow cytometry. Notably, the absence of markers such as CD45, CD11b, Flk-1, CD31, and CD34 confirms their non-hematopoietic and non-endothelial identity. OP9 cells can expand mouse mesenchymal stem cells (MSCs) in vitro, facilitating the study of MSCs' biological features. Their mesenchymal potential has been confirmed through successful differentiation when cultured in an adipogenic medium. When co-cultured with ES cells, OP9 cells enhance the hematopoietic supportive capacity of hemogenic precursors and progeny. Co-culturing ESC with OP9 cells triggers a cascade of events, ultimately leading to the development of various lineages of blood cells. This enables the study of hematopoietic cell differentiation from MSCs, including B cell lineages, erythrocytes, lymphocytes, and megakaryocytes. Furthermore, OP9 cells promote the production of hematopoietic progenitors when multipotent germline stem cells (mGS) are differentiated on their surface. In addition, OP9 cells have demonstrated their ability to facilitate the differentiation of MSCs into osteocytes, chondrocytes, myocytes, tenocytes, and adipocytes. Their migration can be prompted by using growth factors such as bFGF, IGF-1, IL-3, PDGF-BB, TGF-β1, and TGF-β3. However, while OP9 cells offer great promise, there are certain limitations to consider. Since OP9 cells are not an immortalized cell line, the primary cell culture of OP9 cells can only be utilized for short-term, small-scale projects, as it has a limited lifespan of approximately one month.
Organism	Mouse
Tissue	Bone marrow, stroma
Synonyms	OP-9

Age	Embryo
Morphology	Fibroblast-like
Growth properties	Adherent

Citation	OP9 (Cytion catalog number 305174)
Biosafety level	1

Culture Medium	Alpha MEM
Medium supplements	10% FBS, w: 2.0 mM stable Glutamine, w/o: Ribonucleosides, w/o: Deoxyribonucleosides, w: 1.0 mM Sodium pyruvate, w: 2.2g/L NaHCO3
Passaging solution	Accutase
Subculturing	Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.
Split ratio	1:2 to 1:4
Fluid renewal	2 to 3 times per week
Freeze medium	CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100)
Handling of cryopreserved cultures	The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section.
Handling of proliferating cultures	One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 degree Celsius for a minimum of 24 hours.

Sterility	Mycoplasma contaminations are ruled out through PCR-based and luminescence-based mycoplasma assays. The absence of bacterial, fungal or yeast contamination is controlled through daily visual cell monitoring.

Contamination-free cells

To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.

Custom projects

Besides genomic DNA, RNA, cell pellets, and cell lysates, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.

Authenticated cells

Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).

HLA alleles

HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.

OP9

General information

Characteristics

Identifiers / Biosafety / Citation

Expression / Mutation

Handling

Quality control / Genetic profile / HLA