Hs 865.Sk Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	Hs 865.Sk is a finite, non-tumorigenic human cell line of normal skin fibroblasts. It was established from skin tissue obtained from a 59-year-old Caucasian female donor and is derived in situ from the dermal tissue of the skin (skin fibroblasts). As a finite cell line, Hs 865.Sk has a limited proliferative capacity and is often used as a normal, non-cancerous fibroblast control in studies of cellular senescence, aging, and dermal biology. It originates from the same individual as the Hs 869.T cell line. STR profiling confirms its unique identity (Amelogenin X), and it serves as a valuable in vitro model for normal human fibroblast research.
Cell Type	Fibroblast
Tissue	Skin
Tissue Status	Apparently normal tissue
Donor Clinical Context	The donor had gastric adenocarcinoma, but the cell culture was established from apparently normal skin rather than from the gastric tumor
Gender	Female
Age	59 years
Ethnicity	White
Morphology	Fibroblast-like
Growth Mode	Adherent
Potential Research Uses	1. Fibroblast morphology and adhesion studies 2. Three-dimensional cell-culture method development 3. Skin-cell and fibroblast-containing co-culture optimization 4. Matrix-interaction and biomaterial compatibility assays 5. Cytotoxicity and cellular-response method development 6. Imaging, staining, and image-analysis workflow optimization 7. Gene-expression and protein-analysis method development 8. Comparative studies using independently authenticated fibroblast controls
Characteristics
Non-Tumor Tissue Origin	The sampled tissue was described as apparently normal skin even though the donor had gastric adenocarcinoma
Mycoplasma Test	Negative
Culture Conditions and Handling
Culture Temperature	37°C
Culture Surface	Standard tissue-culture vessels support adherent growth. No coating requirement is specified for routine propagation.
Medium Renewal	Every 2–3 days
Dissociation Reagent	0.25% trypsin with 0.03% EDTA
Recommended Split Ratio	1:2 to 1:3
Passage Timing	Passage before the monolayer becomes persistently overconfluent. A 70–90% confluent range is a practical starting point for adherent-cell maintenance, but the optimal threshold should be verified from working-stock morphology and recovery.
Subculturing	1. Remove the spent medium. 2. Rinse the cell layer with 0.25% trypsin and 0.03% EDTA, then remove the rinse. 3. Add 1–2 mL of the same trypsin-EDTA solution. 4. Hold at room temperature or 37°C until the cells detach, using microscopic observation to avoid excessive exposure. 5. Add fresh complete medium and disperse the cells gently. 6. Transfer the suspension to new vessels at a 1:2 to 1:3 ratio. 7. Return the cultures to the validated 37°C incubation environment and replace medium every 2–3 days.
Detachment Control	Observe the monolayer during trypsinization and neutralize the dissociation reagent promptly with serum-containing complete medium after detachment. Excessive enzymatic exposure can reduce attachment and recovery.
Cell Dispersion	Use gentle pipetting to obtain a uniform suspension while minimizing mechanical stress. Persistent clumps can produce uneven seeding and distort cell-count and morphology comparisons.
Thawing	Thaw rapidly with gentle agitation in a 37°C water bath for approximately 2 minutes. Disinfect the vial exterior and transfer the contents aseptically into pre-warmed complete medium.
Cryoprotectant Removal	For a general recovery workflow, dilute the thawed suspension into approximately 9 mL complete medium, centrifuge at about 125 × g for 10 minutes, remove the supernatant, and resuspend the pellet gently. Validate this step against lot-specific recovery behavior.
Cryopreservation Medium	Complete growth medium supplemented with 5% DMSO
Long-Term Storage	Below −130°C, preferably in the vapor phase of liquid nitrogen; −70°C is not suitable for continued storage
Post-Thaw Monitoring	Inspect attachment, fibroblast-like morphology, debris, and medium condition after recovery. Delay experimental use until growth is stable and the culture meets predefined identity and quality criteria.
Biosafety	Handle as human-derived research material under Biosafety Level 1 or the locally assigned equivalent. Apply site-specific risk assessment, aseptic technique, appropriate personal protective equipment, decontamination, and waste procedures.
Handling Notes	Record passage number, seeding density, confluence, split ratio, medium and serum lot, time after seeding, and morphological observations. Clearly distinguish the parental culture from gene-edited, reporter-labeled, selected, or otherwise adapted derivatives.

Cat.No.	Product Name	Price
CSC-RO02646	Cas9 Stable Cell Line - Hs 865.Sk	Inquiry

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* For research use only. Not intended for any clinical use.

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