Types of Growth Factors for Common Cell Types

Growth and attachment factors, natural and recombinant, are used in conjunction with other essential nutrients and hormones to maximize biological stimulation. The optimal conditions and concentrations of growth factors for cell types must be empirically determined. The 5 main types of growth factors used in cell culture are:

EGF (epidermal growth factor), used for culturing epithelial and fibroblast cells
FGF-2 (basic fibroblast growth factor), also known as FGF basic or bFGF and used for maintenance of undifferentiated cells
BDNF (brain-derived neurotrophic factor), for promoting neuronal survival and differentiation
VEGF (vascular endothelial growth factor), used to induce endothelial cell proliferation and angiogenesis
GM-CSF (granulocyte macrophage colony-stimulating factor), for modulating hematopoietic cells

On this page:

Cell line and growth factor examples for cultures
Materials for optimizing media with growth factors
Protocol for optimizing media with growth factors
- Reconstitution and storage of recombinant proteins
- Add growth factors to media and sera

The following table provides a summary of conditions for growth and attachment factor supplementation reported to give maximal stimulation for various cell lines. It can be used to establish initial concentrations prior to optimization; for the best results please refer to your product’s growth factor specifications.

Cell line and growth factor examples for cultures

Cell line	Cell type	Recommended growth factor or attachment factor	Growth factor concentration range (ng/mL)	Medium	Serum supplementation
–	Primary hepatocyte	EGF	10	DMEM	10% FBS
–	Primary human fibroblast	EGF	1 x 10³	MEM Earle's	10% FBS
A431	Epidermal carcinoma	EGF	2.5 x 10⁴	DMEM	10% FBS
A549 [4]	Lung carcinoma	EGF	50	Ham’s F-12K or DMEM	10% FBS
BALB/c 3T3	Mouse fibroblast	EGF	50	DMEM/F-12	–
BHK-21	Baby hamster kidney	FGF-2 (bFGF)/ hs FGF-2	3	DMEM/F-12	5% Calf
C6 [5,6]	Glial cells	BDNF	–	DMEM, high glucose	2.5% FBS
C8-D1A	Astrocyte	BDNF	–	DMEM	10% FBS
Daudi [7]	Burkitt's Lymphoma	GM-CSF	–	RPMI 1640	10% FBS
FDC-P1 [8,9]	Mouse bone marrow	GM-CSF	–	DMEM	10% FBS
GH3	Rat neuroblastoma	FGF-2 (bFGF)/ hs FGF-2	1	F-12	–
H9 (ESC)	Embryonic stem cells	FGF-2 (bFGF)/ hs FGF-2	4	Gibco StemFlex Medium and supplement	KnockOut Serum Replacement
HAEC [10,11]	Aortic endothelial cells	VEGF	–	Human Large Vessel Endothelial Cell Basal Medium	Large Vessel Endothelial Supplement
HCAEC [12]	Human coronary artery endothelial cells	VEGF	–	Human Large Vessel Endothelial Cell Basal Medium	Large Vessel Endothelial Supplement
HeLa	Human cervical carcinoma	EGF	2.5 x 10⁴	DMEM	10% FBS
HF	Human foreskin fibroblast	EGF	2	DMEM	10% FBS
HL-60 [13,14]	Leukemia	GM-CSF	0.01–100 ng/mL	IMDM	20% FBS
HPAEC [10,15,16]	Pulmonary artery endothelial cells	VEGF	–	Human Large Vessel Endothelial Cell Basal Medium	2% FBS
HUVEC [7]	Human umbilical vein endothelial cells	VEGF	–	Ham’s F-12K	10% FBS
iPSC [17]	Induced pluripotent stem cells	FGF-2 (bFGF)/ hs FGF-2	–	Gibco Essential 8 Medium	RevitaCell Supplement (100X)
MCF 7	Human mammary carcinoma	EGF	100	DMEM	2% FBS
MKN-7	Human adenocarcinoma	EGF	50	DMEM	8% FBS
Neuro-2A	Neuroblastoma	BDNF	–	DMEM	10% FBS
NRK-49F	Rat fibroblast	EGF	50	DMEM/F-12	–
SH-SY5Y [5]	Neuroblastoma	BDNF	–	DMEM	10% FBS
TF-1 [5]	Erythroleukemia	GM-CSF	2	RPMI 1640	10% FBS
THP-1 [18,19]	Monocytic leukemia	GM-CSF	–	RPMI 1640	10% FBS
TM-4	Mouse testes epithelium	EGF	3	DMEM/F-12	–
U-87 MG	Glioblastoma	BDNF	–	DMEM	10% FBS
U-937 [18,19]	Monocytic leukemia	GM-CSF	–	RPMI 1640	10% FBS
WB	Rat hepatic epithelium	EGF	10	Richter's MEM	10% FBS

Don’t see your cell line? Our Cell Culture Select Tool features over 150 common cell types for a variety of applications. Simply enter in your chosen cell line and see results for the recommended medium, supplements, and cryopreservation environment—then, when you’re ready, add them to your cart for simplified shopping.

Materials for optimizing culture media with growth factors

The following list of materials is generic and should be used as a guide to collect the necessary materials to complete your experiment. For a complete list of materials needed, please refer to the product-specific instructions for your cell line.

Laminar flow hood
70% ethanol
Personal protective equipment (PPE)
Vial(s) of your selected cell line
The appropriate growth supplements, serum, and basal medium to make your desired complete growth medium
Antibiotics such as Penicillin-Streptomycin (optional)
Cell culture plates, dishes, and flasks
Sterile pipettes and pipette tips
Sterile conical tubes

Protocol for optimizing culture media with growth factors

The following protocol does not specify the appropriate concentration of working medium, serum, reconstitution buffer, or types of growth factors needed to mix a complete growth medium. For specific instructions related to media preparation for your chosen cell lines, download the Product Sheet and Certificate of Analysis (CoA) to obtain the necessary information before beginning your experiment. Failure to do so will result in incorrect concentration or possible damage to the products. Always work in aseptic conditions to avoid contamination.

Reconstitution and storage of recombinant proteins

Without opening the vial containing the lyophilized growth factor, start by centrifuging the vial at approximately 13,000 rpm for 30–60 seconds.
Reconstitute the growth factor by dissolving the lyophilized protein in the appropriate solvent and concentration according to the lot-specific CoA. Do not add carrier to the recombinant protein’s initial reconstitution solution unless otherwise specified on the CoA. (Most proteins can be reconstituted with the addition of sterile distilled water at a concentration of 0.1 to 1.0 mg/mL. For example, for 100 µg of protein, the amount of water that should be added should be between 100 µL and 1 mL, resulting in a protein solution with a concentration between 1 mg/mL and 0.1 mg/mL. However, the product data sheet or COA will indicate when a diluent other than water is required. Recommended solutions, carrier protein concentrations, and extended storage conditions can also be found within these documents.)
Mix carefully by gently pipetting up and down or put the vial cap back on and invert the vial gently a few times. Do not vortex after reconstitution.
Spin down the vial for a few seconds.
Make storage preparations for your proteins:
- The reconstituted protein solution can be stored at 2–8°C for up to one week until you are prepared to use it in your experiment.
- To create aliquots for long-term freezer storage, dilute to working concentration using a buffer or medium containing a carrier (0.1% BSA, 5% HSA, 10% FCS, 5% trehalose 2–8˚C –20°C to –80°C.) Distribute to aliquots and store at –20 to –80°C (for 3–12 months, as specified on the CoA.) Aliquots should not have a concentration of less than 1 µg/mL and/or a volume of less than 10 μL.

Add growth factors to media and sera

In a sterile laminar flow hood, pipette the required volume of basal growth media into a sterile conical tube or bottle. Ensure all reagents and media are pre-warmed to the appropriate temperature before use.
If required, add serum to the basal growth media to achieve the desired final concentration (typically 10% for fetal bovine serum).
Mix gently by swirling or inverting the tube/bottle to ensure thorough mixing.
Thaw the aliquots of growth factors on ice if they are frozen. Avoid repeated freeze-thaw cycles.
Prepare the required concentration of each growth factor according to the specific needs of your cell line. This typically involves diluting the stock solutions to the working concentrations.
Using sterile pipettes, add the appropriate volume of each growth factor to the basal media and serum. The final concentrations will depend on the specific requirements of your cell line. See the table above for starting concentrations of growth factors for common cell types. Mix gently by swirling or inverting the tube/bottle to ensure thorough mixing.
Label the bottle with the contents and date of preparation.
If not used immediately, store the complete growth medium with growth factors at 4°C and use within 1–2 weeks.

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Useful Numbers for Cell Culture