Human Platelet Lysate FAQ

Most Frequent Questions and Answers About Human Platelet Lysate

*does not contain any heparins (these are added at the point of use). **contains residues of porcine-derived heparin. ᵻ contains residues of fully synthetic heparin.

PRODUCTKEY BENEFITSPACK SIZES AVAILABLEFIBRINOGEN-DEPLETED

REQUIRES HEPARIN?

ANIMAL-COMPONENT FREE
Human Platelet Lysate SolutionAnimal serum-free; most cost effective option25 mL, 100 mL & 500 mLNo

No

Yes*
Human Platelet Lysate -FDAnimal serum-free; fibrinogen-depleted10 mL, 50 mL, 100 mL & 500 mLYes

No

No**
Human Platelet Lysate-XFAnimal component-free; fibrinogen-depleted10 mL, 50 mL, 100mL & 500 mLYes

No

YesƗ
Human Platelet Lysate – CustomizedAnimal serum-free; customisable; small scale process developmentCustomisableNo

Yes

Yes*

hPL is derived from human donor platelets collected from healthy consented volunteer donors at certified blood banks. All individual donors and complete batches are fully tested for pathogens according to 21CFR610.

Both Heparin and Xeno-Free Heparin are shipped at ambient temperature. Store at +4°C on arrival.

All of our hPL products are derived from human platelets and therefore are naturally free form other animal derived material. No animal serum is added during the manufacture of these products.

For a product to be animal-component free, no products can be added to the product during the manufacturing process that are derived from another species. For an animal-component free option (i.e. free from non-human species), either choose Human Platelet Lysate Solution Standard with Xeno-free heparin, or Human Platelet Lysate Solution-XF (which is manufactured using Xeno-free heparin, a fully synthetic heparin product).

Human Platelet Lysate (hPL) can be used at a variety of concentrations, from 10% in basal media to as little as 2.5%.  The table below indicates the concentrations of hPL used for a variety of different cell types and applications.

Human Platelet Lysate (hPL) is a cell culture growth supplement obtained from human platelets. hPL contains abundant human growth factors including PDGF, bFGF, EGF, TGF-beta1, VEGF, other cytokines and proteins which yield superior results when culturing many types of human cells, particularly human MSC, regardless of their tissue source.

Serum (traditionally FBS or Human) is the supernatant of clotted blood, hPL is manufactured from lysed platelets.

Performance: The future of cell therapy is to be xenogeneic-free and therefore human platelet lysate offers xenogeneic free/serum substitutes for replacing FBS and improving cell culture performance.

Our ‘Xeno-Free’ products are non-xenogeneic and animal serum-free. i.e. they do not contain material from species other than human origin.

Materials that are described as being ‘Xenogenic’ are derived from a different species and therefore are not genetically or immunologically compatible. This is very important for cell therapy and clinical applications.

For both Human Platelet Lysate-XF (Xeno-Free) and Human Platelet Lysate-FD (Fibrinogen Depleted), a fibrinogen depletion step is obtained during the manufacturing process in order to remove all fibrins. In addition, heparins are added during the manufacturing process. This means that you do not need to add Heparin while preparing the cell culture mix.

For Human Platelet Lysate FD, the heparin used in manufacturing is derived from porcine intestine (and therefore not xeno-free).

For Human Platelet Lysate XF, the heparin used is a xeno-free version, and is fully synthetic, offering a fully xeno-free end product.

Research Grade material is manufactured to meet the needs of basic research and development. This material supports the in vitro propagation and maintenance of various human cell types. In addition, it provides a cost effective alternative to using serum-free media. Material is supplied in validated PETG NALEGENE bottles (25 mL, 100 mL and 500 mL).

GMP Grade material is manufactured under GMP-controlled conditions for use in GMP compliant cell culture SOPs. This material is provided in cryo-bags (100 mL) or in validated PETG NALEGENE bottles (100 mL and 500 mL).

GMP Climical Grade material is manufactured and gamma irradiated under GMP conditions to comply with the high safety guidelines for clinical applications. Due to pathogen-reduction, GMP Clinical Grade hPL provides a high safety level for expansion of different human cell types (particularly adult stem cells, e.g. MSCs) for clinical and therapeutic applications.

Only the Human Platelet Lysate Standard and Customised products require the addition of heparin. There are two types of heparin available:

  1. Heparin – Standard animal-derived heparin (5,000 U/mL) – HSUP-001M.
  2. Xeno-Free Heparin – Animal component-free heparin, fully synthetic and preservative-free (12.5 mg/mL) – HSUP-XF-001M

Heparin is an anti-coagulant that prevents the fibrinogen in human platelet lysate from clotting in the cell culture medium during cell expansion. Both Human Platelet Lysate XF and Human Platelet Lysate FD are fibrinogen-depleted and so do not require the addition of heparin in the cell culture media.

Human Mesenchymal Stromal Cells from Bone Marrow (hMSC-BM)

Human Mesenchymal Stromal Cells from Umbilical Cord (hMSC-UC)

Human Mesenchymal Stromal Cells from Adipose Tissue (hMSC-AT)

Human Dermal Fibroblasts (HDF)

In addition, see FAQ ‘Which cell types has hPL been shown to work in?’, for a more detailed table of other cell types that hPL has been shown to work with.

No. hPL is for in vitro experimental and research use only (RUO) and the product is not intended for human or animal diagnostic or therapeutic uses.

Each batch of hPL is tested to ensure the absence of bacteria, fungi, yeast, mycoplasma (M. fermentans, M. arginini, M. orale, M. hyorhinis, M. salivarium, M. hominis, M. pneumoniae, Acholeplasma laidlawii, M. synoviae and Ureaplasma species). Batches are also tested for the ability to support growth of specific cell lines. In addition, each batch is also tested for standard parameters. These include endotoxin, pH, osmolality and total protein. Details for each batch are supplied on the Certificate of Analysis (COA).

Our hPL products are manufactured from platelet units obtained from healthy blood donors who have been tested and found negative for HBsAg, HIV I/II antibody, CMV, HCV- and HBC-Antibody, HIV-, HCV-, HBV-, HAV-, PVB19-NAT, Treponema pallidum and Syphilis.

hPL is aseptically processed. Each batch of hPL is tested for the absence of bacteria, fungi, yeast, and mycoplasma. Batches of hPL are also sterile filtered. Results of microbe testing of each batch are supplied in the COA. However, please note that sterility is not guaranteed after opening. The use of aseptic techniques during handling is recommended.

A gamma-irradiated option (GMP-Clinical grade) for the purpose of virus-inactivation is available for all hPL products. NOTE: our gamma-irradiated products are manufactured in the US.

Human Platelet Lysate Standard – requires the addition of heparin to the final cell culture medium. Choose from a standard animal derived heparin (porcine), or a xeno-free heparin (fully synthetic and preservative free). For human ex-vivo tissue and cell culture processing applications.

Human Platelet Lysate FD (Fibrinogen Depleted) – A fibrinogen-depleted hPL that does not require the addition of heparin. However, it may contain residues of heparin. For human ex-vivo tissue and cell culture processing applications.

Human Platelet Lysate XF (Xeno-Free) – A completely xeno-free fibrinogen-depleted hPL. Does not require the addition of heparin, however it may contain residues of synthetic, xeno-free heparin. For human ex-vivo tissue and cell culture processing applications.

Human Platelet Lysate-i – A gamma irradiated GMP-Clinical grade culture supplement. Each of the above three options can be supplied gamma irradiated for regulatory compliance. Suitable for cell therapy purposes.

Human Platelet Lysate Customised – A custom-manufactured human platelet lysate solution that requires the addition of heparin to the final cell culture medium. Designed for the small-scale process development needs of biopharmaceutical life cycles, protein therapeutics, biologics, vaccine expression or small-volume testing prior to GMP scale-up.

Research Grade: 25 mL, 100 mL and 500 mL NALGENE bottles.

GMP Grade: 100 mL cryo-bags and 100 mL NALGENE bottles and 500 mL NALGENE bottles.

GMP-Clinical Grade (gamma irradiated): 50 mL, 100 mL and 500 mL NALGENE bottles.

Yes. Samples of hPL are available for testing prior to selection of a suitable batch. Typical sample size is 25 mL and reservations are held for a period of four weeks, pending evaluation.

hPL has a shelf life of 24 months from the date of manufacture, provided they are stored appropriately. We would recommend enquiring about the shelf life of each available batch if it is important to have a long shelf life following purchase.

The recommended storage is -20°C or below.

hPL products can be storage long term at -80°C.

Thaw frozen hPL product overnight at 4°C or for 1 hour in a 37°C water bath before use. Once hPL product is thawed, it is recommended to fully use it or to aliquot and re-freeze samples of unused hPL at -20°C.  Completed medium preparation (e.g. 5% hPL) can be stored at 4°C and is stable for approximately four weeks.

It is recommended to avoid freeze-thaw cycles as this can lead to an increase in the formation of insoluble particles.

All biological material should be handled as potentially infectious. It is essential that universal precautions should be employed when handling hPL.

Despite all testing, proper safety precautions for potentially infectious agents must be taken. All human blood products should be handled in accordance with currently acceptable bio-safety practices and guidelines for the prevention of blood borne viral infections.

hPL will be shipped frozen, by overnight courier in polystyrene moulded boxes with dry ice. This will ensure that the material arrives frozen in good condition.

The presence of particulate matter is a result of fibrinogen converting to insoluble fibrin resulting from freezing and thawing human platelet lysate. This particulate matter does not indicate contamination of any sort, and does not have any negative impact on cell culture performance. To avoid particulate matter from forming, we suggest minimal freeze / thaw cycles when using our products.

Should insoluble particles appear in thawed hPL we recommend removing particles by centrifuging at 3,400 x g for 3-5 minutes.

If clotting or insoluble particles appears in the final medium, we recommended filtering the complete culture medium after diluting in the basal medium to remove insoluble particles. Filtering the completed medium (e.g. 5%), after hPL is diluted in the basal medium, will not affect supplemented cell culture performance. However, filtering is NOT recommended for 100% concentrate hPL as this may reduce cell culture performance.

Technically and procedurally there is no any difference between our research and GMP Grade products. The difference is in the additional documentation supplied for the manufacturing process of the GMP Grade material.

Based on our testing we can see minimal differences between both products in terms of performance. The xeno-free products were developed to provide the same levels of performance but in a xeno-free environment.

  1. Thaw Human Platelet Lysate Solution overnight at 4°C or for 1 hour in a 37°C water bath.
  2. Prepare complete MSC culture medium by adding uman Platelet Lysate Solution to basal medium (i.e. Dulbecco’s Modified Eagles Medium-Low Glucose; DMEM-LG) with 2 mM L-glutamine and 100 U/mL Penicillin/streptomycin as final concentration.
  3. hPL shows optimal growth of MSC at 5% (v/v). However, for higher cell proliferation rates, it is recommended to use 10% (v/v) of hPL.
  4. We recommend seeding MSCs at approximately 3×103 ~ 6×103 per cm2.
  5. For Human Platelet Lysate Solution ONLY, to avoid coagulation of the complete MSC culture medium, Heparin should be added at a final concentration of 2 U/mL Heparin (cat.no. HSUP-001M) or 0.024 mg/mL Xeno-Free Heparin (cat. no. HSUPXF-001M).

NOTE: Human Platelet Lysate FD and Human Platelet Lysate XF DO NOT require the addition of heparin as they are fibrinogen depleted products.

  1. Complete MSC culture medium can be stored at 4°C and is stable for approximately four weeks.

From a technical viewpoint, the application of hPL, Human serum (HS) and FBS are very similar. hPL differs to sera products due to the increased volume of cytokines and growth factors contained within the product affecting the performance levels.

We would recommend the following paper by Bieback et al. 2009 that demonstrates the comparative performance difference between hPL, HS and FBS.

Bieback, K., et al. (2009) Human alternatives to fetal bovine serum for the expansion of mesenchymal stromal cells from bone marrow. Stem Cell, 27(9):2331-2341.

SPECIESCELL TYPECELL NAMEDESCRIPTION

CONCENTRATION USED (%)

BovinePrimary CellCECCorneal Endothelial Cells

2.5 – 10%

HamsterCell LineCHOChinese Hamster Ovary epithelial

Unknown

HumanCell LineHEK-293Human Embryonic Kidney 293 cells

1 – 10%

HumanCell LineHeLaHeLa Cervical cancer cells

10%

HumanCell LineHFFF2HFFF2 Dermal fibroblasts

5 – 20%

HumanCell LineHL-60Human leukaemia cell lines

20%

HumanCell LineHUVECHuman umbilical vein endothelial cells

5 – 20%

HumanCell LineJURKATHuman leukaemia cell lines

10%

HumanCell LineK562Myelogenous leukaemia

10%

HumanCell LineKG-1Human leukaemia cell lines

20%

HumanCell LineMCF-7Human continuous tumour

10%

HumanCell LineNCTC 2544Dermal keratinocytes

5%

HumanCell LineNCTC 2544Dermal keratinocytes

5 – 20%

HumanCell LineA-549Human continuous tumour

5%

HumanCell LineCaco-2Human continuous tumour

5%

HumanCell LineHaCaTKeratinocyte cell line from adult human skin

5%

HumanCell LineHEK-293Human Embryonic Kidney 293 cells

Unknown

HumanCell LineHGF-1Human Gingiva Fibroblasts

5%

HumanCell LineMCF-7Human continuous tumour

10%

HumanCell LineU-251 MGHuman continuous tumour

5%

MouseCell LineC2C12Myoblastoma Cells

1 – 20%

MouseCell LineMMT 060562Mouse mammary tumour

Unknown

MouseCell LineNeuro-2aNeuroblastoma cell line

Unknown

MouseCell LineNIH/3T3Mouse Embryonic Fibroblasts

1 – 10%

MouseCell LineRAGMouse adenocarcinoma cell line

Unknown

RabbitCell LineSIRCStatens Seruminstitut Rabbit Cornea

1 – 10%

RatPrimary CellMSCMesenchymal Stem Cells

2.5 – 5%

RatCell LineR2CTestis from rat

Unknown

The above list represent cell lines and conditions that have been stated in published literature or from customer testimonials. We would recommend testing your cells with a titration range to determine the ideal conditions for your particular cells and application.