Primary Lung Fibroblast, Normal, Human (HLF) (ATCC® PCS-201-013)

Organism: Homo sapiens, human  /  Tissue: Lung  /  Cell Type: Fibroblast

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Organism Homo sapiens, human
Tissue Lung
Cell Type Fibroblast
Morphology spindle-shaped
Growth Properties adherent
Biosafety Level 1 [These primary cells are not known to harbor an agent recognized to cause disease in healthy adult humans. Handle as a potentially biohazardous material under at least Biosafety Level 1 containment. Cells derived from primate lymphoid tissue may fall under the regulations of 29 CFR 1910.1030 Bloodborne Pathogens. 

ATCC recommends that appropriate safety procedures be used when handling all primary cells and cell lines, especially those derived from human or other primate material. Detailed discussions of laboratory safety procedures are provided in Laboratory Safety: Principles and Practice, 2nd ed. (ASM Press, Washington, DC) (Fleming et al., 1995) and Caputo, J.L. Biosafety procedures in cell culture. (1988) J. Tissue Culture Methods 11:223. Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at] 

Human Material Precaution & nbsp;

All tissues used for isolation are obtained under informed consent and conform to HIPAA standards to protect the privacy of the donor & rsquo;s personal health information. It is best to use caution when handling any human cells. We recommend that all human cells be accorded the same level of biosafety consideration as cells known to carry HIV. With infectious virus assays or viral antigen assays, even a negative test result may leave open the possible existence of a latent viral genome.

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease normal
Age lot-specific
Gender lot-specific
Ethnicity lot-specific
Applications An ideal culture model for the study or development of a potential diagnostic method for the early detection of lung cancer cells, reconstruction studies, and advancement of cancer research.
Product Format frozen 1.0 mL
Storage Conditions -130°C or below
Images Lung Fibroblast TE 7

Lung fibroblast cells (HLFs) are cryopreserved at P2 and marketed as secondary cells (cells have been isolated, plated, and expanded in culture vessels twice prior to cryopreservation) to ensure the highest viability and proliferation efficiency.

Complete Growth Medium
  1. Obtain one vial of Primary Lung fibroblast cells ; Normal, Human (ATCC PCS-201-013) from the liquid nitrogen tank; make sure that the caps of all components are tight.
  2. Thaw the components of Fibroblast Growth Kit-Low serum ATCC® PCS-201-041™just prior to adding them to the Fibroblast Basal Medium ATCC® PCS-201-030™ .
  3. Obtain one bottle of Fibroblast Basal Medium ATCC® PCS-201-030™ from cold storage.
  4. Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying them with 70% ethanol.
  5. Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component, as indicated in Table 1, into the bottle of basal medium using a separate sterile pipette for each transfer.

Table 1. Fibroblast Growth Kit-Low Serum Components



    Final Concentration

    rh FGF b

    0.5 mL

    5 ng/mL


    18.75 mL

    7.5 mM

    Ascorbic acid

    0.5 mL

    50 µg/mL

    Hydrocortisone Hemisuccinate

    0.5 mL

    1 µg/mL

    rh Insulin

    0.5 mL

    5 µg/mL

    Fetal Bovine Serum

    10.0 mL


  1. Tightly cap the bottle of complete growth medium and swirl the contents gently to assure a homogeneous solution. Do not shake forcefully to avoid foaming. Label and date the bottle.
  2. Complete growth media should be stored in the dark at 2°C to 8°C (do not freeze). When stored under these conditions, complete growth media is stable for 30 days.

  1. Passage normal lung fibroblast cells when culture has reached approximately 80% to 90% confluence, and are actively proliferating.
  2. Warm both the Trypsin-EDTA for Primary Cells (ATCC PCS-999-003) and the Trypsin Neutralizing Solution (ATCC PCS-999-004) to room temperature prior to dissociation. Warm complete growth medium to 37°C prior to use with the cells.
  3. For each flask, carefully aspirate the spent media without disturbing the monolayer.
  4. Briefly rinse the cell layer with 3 to 5 mL DPBS (ATCC 30-2200) to remove residual traces of serum and then aspirate and discard the DPBS.
  5. Add pre-warmed trypsin-EDTA solution (2 to 3 mL for every 25 cm2) to each flask.
  6. Gently rock each flask to ensure complete coverage of the trypsin-EDTA solution over the cells.
  7. Observe the cells under the microscope. When the cells pull away from each other and round up (typically within 3 to 5 minutes), remove the flask from the microscope and gently tap it from several sides to promote detachment of the cells from the flask surface.
  8. When the majority of cells are detached, quickly add an equal volume of Trypsin Neutralizing Solution (ATCC PCS-999-004) to each flask. Gently pipette or swirl the culture to ensure all of the trypsin-EDTA solution has been neutralized.
  9. Transfer the dissociated cells to a sterile centrifuge tube and set aside while processing any remaining cells in the flask.
  10. Add 3 to 5 mL Trypsin Neutralizing Solution (ATCC PCS-999-004) to the flask to collect any remaining dissociated cells. Transfer remaining cells into the centrifuge tube.
  11. Repeat steps 10 as needed until all cells have been collected from the flask.
  12. Centrifuge the cells at 150 x g for 3 to 5 minutes.
  13. Carefully aspirate the neutralized dissociation solution from the cell pellet and re-suspend the cells in 5 to 8 mL fresh, pre-warmed, complete growth medium.
  14. Count the cells and seed new flasks at a density of 2,500 to 5,000 cells per cm2.
  15. Place freshly seeded flasks in a 37°C, 5% CO2, incubator for at least 24 to 48 hours before processing the cells further. Refer to Maintenance for guidelines on feeding.
Volume 1.0 mL
Sterility Tests Bacteria and Yeast: Negative
Mycoplasma: Negative
Viral Testing Hepatitis B: Negative
Hepatitis C: Negative
HIV: Negative
Viability ≥ 70% when thawed from cryopreservation
C of A
Certificate of Analysis
Certificate of Analysis
Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation