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Fibroblast Growth Kit-Low serum

PCS-201-041

Fibroblast Growth Kit–Serum-Free (ATCC PCS-201-040) and Fibroblast Growth Kit–Low Serum (ATCC PCS-201-041) each contain components that when added to Fibroblast Basal Medium (ATCC PCS-201-030) create a complete ATCC Primary Cell Solutions™ culture environment for fibroblasts (e.g., Dermal Fibroblasts, Normal, Human Neonatal [ATCC PCS-201-010], Dermal Fibroblasts, Normal, Human Neonatal, Mitomycin C Treated [ATCC PCS-201-010]).

Product category
Reagents
Comments

An upper limit for population doublings has not been established for primary fibroblasts when cultured with Fibroblast Basal Medium supplemented with Fibroblast Growth Kit, Low serum (2% FBS).
The final concentration for each component is as follows:

  • L-glutamine: 7.5 mM
  • rh FGF basic: 5 ng/mL
  • rh Insulin: 5 µg/mL
  • Hydrocortisone: 1 µg/mL
  • Ascorbic acid: 50 µg/mL
  • Fetal bovine serum: 2%
Applications
Cell growth and viability
Product format
Frozen
Shipping information
1 kit
Storage conditions
-20°C or colder, -70°C for long-term storage
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Documentation

Biosafety Icon BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Detailed product information

General

Specific applications

The kit contains components that when added to the fibroblst basal medium ATCC PCS-201-030 create a complete culture environment for fibroblasts. Use of this kit will support a faster rate of proliferation compared to when Fibroblast Growth Kit, Serum-free (ATCC PCS-201-040) is used.

Your experimental design will dictate which Fibroblast Growth Kit should be used. The use of Fibroblast Growth Kit–Serum-Free creates a completely defined medium for the serum-free culture of human fibroblasts. The rate of proliferation is equal to or greater than media supplemented using FBS (at concentrations ranging from 2% to 10%) through 10 population doublings. (Proliferation rate may slow under serum-free conditions after 10 population doublings.) Under these conditions, serum-free human feeder cell layers can be established for stem cell culture, or as a model system to study wound healing, skin aging, gene delivery, toxicology and basic questions of cell biology. 

Use of the Fibroblast Growth Kit–Low Serum (ATCC PCS-100-041) will support a faster rate of proliferation compared to Fibroblast Growth Kit–Serum-Free (ATCC PCS-100-040. An upper limit for population doublings has not been established for primary fibroblasts when cultured with Fibroblast Basal Medium supplemented with Fibroblast Growth Kit–Low Serum (2% FBS). Under these conditions, primary fibroblast cultures have been maintained for 30+ population doublings, performing well when used as feeder layers to support human ES cell cultures.

Handling information

Handling procedure
  1. Obtain one growth kit from the freezer; make sure that the caps of all components are tight.
  2. Thaw the components of the growth kit just prior to adding to the basal medium. It is necessary to warm the L-glutamine component in a 37°C water bath, and shake to dissolve any precipitates, prior to adding to the basal medium.
  3. Obtain one bottle of Fibroblast Basal Medium (480 mL) 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 volume of each growth kit component, as indicated in Table 1 or 2, to the bottle of basal medium using a separate sterile pipette for each transfer.
  6. Table 1. If using the Fibroblast Growth Kit–Serum-Free (ATCC® PCS-201-040), add the indicated volume for each component in the order shown.

    Component

    Volume

    Final Concentration

    L-glutamine

    18.75 mL

    7.5 mM

    Hydrocortisone Hemisuccinate

    0.5 mL

    1 μg/mL

    HLL Supplement

    1.25 mL

    HSA 500 μg/mL

    Linoleic Acid 0.6 μM

    Lecithin 0.6 μg/mL

    rh FGF basic

    0.5 mL

    5 ng/mL

    rh EGF / TGF  β-1 Supplement

    0.5 mL

    5 ng/mL

    30 pg/mL

    rh Insulin

    0.5 mL

    5 μg/mL

    Ascorbic acid

    0.5 mL

    50 μg/mL

    Table 2. If using the Fibroblast Growth Kit–Low

    Serum (ATCC® PCS-201-041), add the indicated volume for each of the following components:

    Component

    Volume

    Final Concentration

    rh FGF basic

    0.5 mL

    5 ng/mL

    L-glutamine

    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

    2%

    Antimicrobials and phenol red are not required for proliferation but may be added if desired. The recommended volume of each optional component to be added to the complete growth media is summarized in Table 3.

    Table 3. Addition of Antimicrobials/Antimycotics and Phenol Red (Optional)

    Component

    Volume

    Final Concentration

    Gentamicin-Amphotericin B Solution

    0.5 mL

    Gentamicin: 10 µg/mL

    Amphotericin B: 0.25 µg/mL

    Penicillin-Streptomycin-Amphotericin B Solution

    0.5 mL

    Penicillin: 10 Units/mL

    Streptomycin: 10 µg/mL

    Amphotericin B: 25 ng/mL

    Phenol Red

    0.5 mL

    33 µM

     

  7. 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.
  8. 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
Cryopreservation
1. Check all containers for leakage or breakage.
2. Store the growth kit(s) at either -20°C in a freezer that is not self-defrosting or at -70°C for long-term storage. If thawed upon arrival, the Growth Kit can be stored at 2°C to 8°C and added to the basal medium within 72 hours of receipt.

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Functional tests
Rate of proliferation and morphology of fibroblasts.
*A Certificate of Analysis (COA) is available upon request for each lot of Fibroblast Growth Kit.

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

References

Frequently Asked Questions

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