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INTENDED USE
This product is for research use only (RUO). Not for use in diagnostic or therapeutic procedures.

IMPORTANT USER NOTE
These cells exhibit biological characteristics associated with the obese donor phenotype, which may impact their growth, differentiation, and metabolic behavior compared to cells from lean donors. It is the responsibility of the user to determine the suitability of this product for their specific application. Proper training in the handling of human-derived materials is required.

PRODUCT DESCRIPTION

Regeneration Biology offers Human Adipose-Derived Stem Cells (ADSCs) from obese donors, isolated from the subcutaneous adipose tissue of donors with a clinically defined Body Mass Index (BMI) of 30 kg/m² or greater. These multipotent mesenchymal stem cells are cryopreserved at passage 1 (P1) to ensure optimal viability and functionality.

ADSCs from an obese background provide a critical tool for studying the pathophysiology of obesity, metabolic syndrome, and their related complications. These cells exhibit a distinct molecular and secretory profile compared to lean-derived ADSCs, including altered adipokine secretion, increased inflammatory marker expression, and potential differences in differentiation capacity. They are essential for modeling disease mechanisms, screening therapeutic compounds, and investigating regenerative medicine in a metabolically challenged context.

KEY FEATURES & BENEFITS

• Disease-Relevant Model: Sourced from verified obese donors (BMI ≥ 30), providing a physiologically relevant model for obesity research.

• Multipotent Potential: Retain the capacity to differentiate into adipocytes, osteocytes, and chondrocytes under appropriate conditions, allowing for studies on lineage commitment in an obese microenvironment.

• Characterized Immunophenotype: Express classic mesenchymal stem cell surface markers (CD73, CD90, CD105) and lack expression of hematopoietic markers (CD14, CD34, CD45, HLA-DR).

• Functional & Quality Tested: Every lot is tested for viability, sterility, and differentiation potential to ensure performance.

• Comprehensive Donor Data: Supplied with extensive donor information, including BMI, comorbidities (e.g., Type 2 Diabetes, hypertension), and relevant clinical history.

QUALITY CONTROL SPECIFICATIONS

Each lot is tested to meet the following release criteria:

Viability (post-thaw): ≥ 80% (Test Method: Trypan Blue Exclusion)
Cell Yield per Vial: ≥ 1 million or 5 million viable cells (Test Method: Cell Count)
Adherent Growth: Positive for fibroblastic, spindle-shaped morphology (Test Method: Microscopic Evaluation)
Surface Marker Expression: Positive (≥95%): CD73, CD90, CD105; Negative (≤5%): CD14, CD34, CD45, HLA-DR (Test Method: Flow Cytometry)
Trilineage Differentiation*: Positive for Oil Red O (Adipogenesis), Alizarin Red S (Osteogenesis), and Alcian Blue (Chondrogenesis) (Test Method: Functional Staining)
Microbiological Sterility: No growth (Test Method: USP <71>)
Mycoplasma: Negative (Test Method: PCR or Culture Method)

APPLICATIONS

• Obesity & Metabolic Disease Research: Study altered adipokine secretion, chronic inflammation, and insulin resistance in vitro.

• Drug Discovery & Screening: Evaluate efficacy of anti-obesity, insulin-sensitizing, or anti-inflammatory compounds.

• Mechanistic Studies: Investigate adipogenesis, lipid accumulation, and mitochondrial dysfunction in a disease-specific context.

• Co-culture Models: Ideal for establishing systems with hepatocytes, myocytes, or endothelial cells to study inter-tissue crosstalk in metabolic syndrome.

• Regenerative Medicine: Explore the impact of an obese cellular microenvironment on tissue repair and regeneration potential.

HANDLING & STORAGE

• Storage: Store in the vapor phase of liquid nitrogen (below -135°C) immediately upon receipt. Do not store at -80°C for long-term preservation.

• Shipment: Shipped on dry ice. Upon receipt, transfer vials to liquid nitrogen storage immediately.

• Handling: Use personal protective equipment. All materials handling human primary cells should be treated as potentially infectious and handled under Biosafety Level 2 (BSL-2) conditions.

TYPICAL DONOR DEMOGRAPHICS & CLINICAL HISTORY

Donors are carefully selected and characterized. A typical profile includes:

• BMI Range: 30 - 45 kg/m² (Specific BMI provided per lot)

• Age Range: 25 - 65 years

• Sex: Male and Female donors available

• Common Comorbidities: May include Type 2 Diabetes, Hypertension, Dyslipidemia (specific details provided with Certificate of Analysis)

• Tissue Source: Subcutaneous Adipose (Abdominal or Gluteal)

ORDERING INFORMATION

CELL CULTURE PROTOCOL

Thawing of Frozen Cells:

1. Upon receipt of frozen human adipose-derived stem cells (hADSC), it is crucial to immediately thaw and culture the cells to maximize cell viability. Delays in this process can lead to decreased cell health and functionality.

2. To effectively thaw the cells, submerge the vial containing the frozen cells in a preheated water bath at 37°C. Gently agitate the vial for 1-2 minutes to facilitate uniform thawing. Be careful to keep the cap of the vial above the water level to prevent potential contamination from the water bath, which could compromise sterility.

3. Once the cells are fully thawed, transfer the content of the vial into a sterile 15 mL conical tube that contains 5 mL of fresh Adipose-derived Stem Cell Growth Medium (STEM10). This medium is specifically formulated to support hADSC growth and ensure optimal recovery following thawing.

4. Following the transfer, centrifuge the cells at a speed of 1,000 rpm (approximately 220 × g) for 5 minutes at room temperature. This step helps to pellet the cells, facilitating the removal of any cryoprotectant and cellular debris.

5. After centrifugation, carefully aspirate the supernatant without disturbing the cell pellet. Re-suspend the pellet in fresh Adipose-derived Stem Cell Growth Medium, ensuring thorough mixing to maintain cell viability.

6. Transfer the re-suspended cells into either one 100 mm culture dish or one T75 flask, depending on your experimental requirements. Maintain the culture at 37°C in a humidified atmosphere with 5% CO₂. Change the medium every 2-3 days, monitoring the cells until they achieve a density of 70-80% confluence, indicating readiness for subculturing.

Standard Culture Procedure:

1. Once the cells reach a confluence of 70-80%, perform a careful medium change by first aspirating the existing growth medium. Rinse the cells gently with 5 mL of phosphate-buffered saline (PBS) in a T75 flask to remove any residual serum or growth factors that might inhibit trypsinization.

2. Add 3-5 mL of 0.25% Trypsin-EDTA directly to the flask. Incubate the cells at 37°C for approximately 5 minutes. This enzyme mixture will cleave the adhesive proteins that anchor the cells to the culture surface, thereby enabling cell detachment.

3. Once the incubation period is complete, neutralize the trypsin by adding 2-3 volumes of Adipose-derived Stem Cell Growth Medium to ensure the trypsin action is halted and to provide nutrients for cell recovery.

4. Proceed with centrifugation at 1,000 rpm (approximately 220 × g) for 5 minutes. After the centrifugation step, carefully re-suspend the cell pellet in the desired volume of growth medium, ensuring an even distribution of cells.

5. Seed the new culture vessels at a density of 5 × 10³ cells/cm². This seeding density is essential for optimal cell growth and expansion. Continue to change the medium every 2-3 days, monitoring the cells until they reach 70-80% confluence before considering additional passaging.

KEYWORDS

Adipose-derived stem cells (ADSCs), obesity, donor, stem cell biology, regenerative medicine, adipose tissue, multipotent, differentiation, immunomodulation, metabolic syndrome.

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