| Field | Specification |
|---|---|
| Organism | |
| Applications | |
| Tissue | |
| Disease | Swarm rat chondrosarcoma |
| Growth properties | |
| Biosafety level | |
| Product format | Frozen |
| Storage | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Overview
This product is a stable CRISPR knockout cell line in which Acan (Aggrecan) has been disrupted in the RCS (Rat Chondrosarcoma, Swarm strain) cell line background. RCS cells are a well-established model for chondrocyte biology and cartilage extracellular matrix (ECM) studies; the Acan knockout removes the primary cartilage proteoglycan to enable loss-of-function analysis. Cells grow as adherent and suspension cultures and are supplied frozen (BSL-II).
CRISPR Knockout Design
- Target gene: Acan (Aggrecan; rat gene symbol)
- Knockout strategy: CRISPR-Cas9–mediated disruption of the Acan locus with stable integration confirmed by selection
- Parental cell line: RCS — Rat Chondrosarcoma (Swarm rat chondrosarcoma origin); adherent and suspension growth; bone/cartilage tissue model
Gene Background
Aggrecan (ACAN/Acan) is the most abundant proteoglycan in articular cartilage and is essential for the compressive biomechanical properties of the tissue. Structurally, aggrecan consists of a core protein decorated with glycosaminoglycan chains (chondroitin sulfate and keratan sulfate), which associate with hyaluronan and link proteins to form large aggregating complexes in the pericellular and territorial matrix. Aggrecan expression is regulated developmentally and is considered a marker of chondrocyte differentiation. Loss or degradation of aggrecan by aggrecanases (ADAMTS family enzymes) is associated with cartilage degradation in osteoarthritis and other musculoskeletal diseases. The RCS cell line is a commonly used in vitro model that retains chondrocytic characteristics and produces cartilage matrix components.
Cell Culture Specifications
- Growth properties
- Adherent and suspension
- Organism
- Rat (R. norvegicus)
- Tissue origin
- Bone (Swarm rat chondrosarcoma)
- Biosafety level
- BSL-II
- Format
- Frozen; 1×10⁶ cells / 1.0 ml
Knockout is confirmed by the presence of frameshift-inducing INDELs, verified by Sanger Sequencing.
Research Applications
- Cartilage ECM biology: Investigate the role of aggrecan in proteoglycan network assembly, hyaluronan binding, and matrix hydration in the RCS chondrosarcoma model.
- Aggrecanase target validation: Use the Acan KO line as a negative control to distinguish aggrecan-specific from off-target effects of ADAMTS inhibitors.
- Chondrocyte differentiation: Compare chondrocytic marker expression (Col2a1, Sox9, Col10a1) between Acan KO and parental RCS to assess aggrecan's role in differentiation signaling.
- Glycosaminoglycan analysis: Quantify sulfated glycosaminoglycan secretion (by DMMB assay or Alcian blue staining) in KO vs. parental to establish aggrecan's contribution to total GAG output.
Important Notes
- Mixed growth mode: RCS cells can grow in both adherent and suspension states; culture vessel coating may influence growth behavior. Monitor cell attachment and morphology post-thaw.
- Donor/source: Parental RCS cells originate from the Swarm rat chondrosarcoma model (R. norvegicus).
- Viability warranty: Cell viability upon thaw is warranted for 30 days following shipment when handled according to abm's cell handling guidelines.
- Intended use: For laboratory research use only. Not intended for diagnostic, therapeutic, or clinical applications.
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.
- Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.
- Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.
- Incubate the cells at the recommended conditions.
- Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.
- Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.
- Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.
- Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
- Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
What is your warranty or return policy?
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
