Solutions Guide

1. Title

Section Content Example
Título Include a short title descriptive title of the content of the challenge. Challenge name: New production methodology of adult stem cells free of fetal bovine serum.
Solution title:Breeding methodology free of xeno-components that uses platelet extracts for the production of human stem cells.

2. Abstract

Section Content Example
Abstract Expand briefly as an introduction the purpose of the proposed solution, with particular emphasis on the action mechanisms or scientific basis to justify the potential of the solution.

Replacement of fetal bovine serum in cellular breeding environment by components of human origin contained in platelet extracts from blood donations.

The main advantage of these extracts are as follows:

  • easily accessible as they are routinely discarded by blood banks after expiring and not finding a receiver who needs them..
  • high product quality due to tight production regulation.
  • easily controllable and fewer incompatibility problems than animal substitutes, because of its human origin

Our scientific findings suggest that the richness and complexity of growth factors present in these platelet extracts can be just as effective or even of superior efficacy than traditional methodologies.

3. Technical Scientific Description

Section Content Example
Technical Scientific Description Detailed description of the mechanisms of action and scientific principles of the solution. Indicate how the solution relates to the state of the art today. Detail the results obtained from previous experiments and tests. Explain the advantage of this approach compared to other existing methodologies.

Justificación científico-técnica.

Results obtained using human platelet extracts show that certain cell types such as cardiac stem cells and mesenchymal stem cells can be kept stable after a large number of passes (> 10) in a medium containing 5% platelet extract without further optimizations. Likewise, data from experiments indicate that extracts can be activated to release platelet growth factors using various methods. The release of growth factors, collagen, fibrin and platelets is strongly influenced by the method used for activation. Our results indicate that activation of human platelets by thrombin, which is the natural activator in wounds, is optimal for the release of growth factors necessary for cell lineages as cardiac stem cells and stem cells.

Proposed Process for the preparation of platelet extracts

Platelets are prepared in a standard manner from donations collected with anticoagulant and are isolated by an automatic separator which produces a preparation almost free of leukocytes and with platelets in good condition and without activation.

The received concentrates contain the platelet concentrate of four donations of a unit of blood in a volume ranging between 4 and 15 ml at a concentration of ~ 1-5 x 108 per ml. The average initial concentration of platelets in whole blood system in healthy Spanish population is 2.5 x106 per ml, whereby the concentration of the preparations is between 100 and 200 times the initial one.

To minimize the variation between batches 5 units of platelet are mixed containing platelets from 20 different donors. All donations are reviewed according to the Guideline Note for Guidance on Plasma Derived Medicinal Products (CPMP/BWP/269/95 rev 3) to ensure they are free of pathogens.

The new preparations are purified to decrease the contamination of leukocytes either by centrifugation through a Ficoll gradient and / or by filtration with a filter of PALL leucoreductor (PL1BE), whereby a preparation is obtained with < 1 leucocito/1x106 platelets.

Once purified the platelets are resuspended in human serum to obtain a concentration of 1.5x109 platelets per ml. The preparation labeled and frozen at -800 C until use. This protocol produces a platelet concentrate with a high concentration of growth factors important for stem cells: PDGF-BB: 198 ± 9 ng / ml PDGF-AB: 1845 ± 356; TGF-β1: 120 ± 42 ng / ml , VEGF: 955 ± 375 pg / ml EGF: 470 ± 134 pg / ml IGF-1: 131 ± 45 pg / ml; HGF: 85 ± 34 pg / ml.

Platelet activation will occur through human thrombin. The platelets are diluted to a final concentration of 1x106 per ml in DMEM: F12 (basal medium used for culturing the hCMCs and hMSCs). Then, 2 units / ml of human thrombin is added to the suspension and incubated at 37 ° C for 30 min. To remove the fibrin gel that is formed, the preparation gets centrifuged at 1800 revs for 7 min. at 40 ° C and the preparation is ready to be used fresh or after freezing at -80 ° C for up to 3 months.

4. Feasability

Section Content Example
Feasability Explain why the proposed solution is feasible from a science and technology standpoint.

We compared three different methods of activation for the platelet extracts before deciding which one is the one that contributes to increased production of growth factors that optimize the cell culture process.

The three analyzed methodologies are:

  • Freezing and thawing
  • Activation by human thrombin
  • Platelet activation by calcium

Experiments were carried out ensuring that the produced middle in those three cases respect the quality standards set by relevant regulatory agencies so that resulting medium can be approved for cellular drug manufacturing. The obtained results obtained confirm that the activation method with human thrombin is more effective than the other two and that, at the same time,, enables a stem cell proliferation of various lineages is similar to that obtained using methodologies with fetal calf serum.

5. Suitability

Section Content Example
Suitability Explain why the proposed solution meets the specific requirements detailed by the Challenger in this section: Expected results and commercial application.

The results obtained Throughout the experiment indicate that activation by human thrombin used in short term culture and in a 96 well format, that 1% of platelets is sufficient to maintain cell proliferation of stem cells comparable to the culture media obtained using 1% serum. (FBS).

This level cell proliferation leads to the conclusion that this process is an effective substitute for fetal bovine serum in cell culture.

Comparatively the FBS amount needed to produce a therapeutic dose of stem cells (2 x 109 cells) is 1000 ml. When adding the cost of the growth factors that have to supplement the average FBS growth cost to produce a cell dose currently amounts to € ~ 1900.

As a result of these experiments we can conclude that th platelet activation method proposed in this solution would cost approximately € 700

6. Supporting documents

Section Content Example
Supporting documents Include when appropriate reference to papers, designs, graphics, and / or results of experiments that reinforce the potential of the proposed solution and / or justify the scientific-technical principles on which it is based.
Bruinink, A., Tobler, U., Halg, M et al. Effects of serum and serum heat innactivation on human bone derived osteoblast progenitor cells. J Mater Sci Mater Med 15: 497-501 (2004).

7. Documents

The solver can add all those documents he thinks appropriate to reinforce and / or justify your solution: papers, designs, graphics, and / or results of experiments that reinforce the potential of the proposed solution and / or justify the scientific-technical principles on which it is based.


Comparative graphs of cell proliferation / viability HCSC in presence of 0% FBS, 1% FBS serum and increasing concentrations of platelets (0.5, 1 and 5%).

8. Legal Challenge Conditions

The Challenge Specific Agreement (CSA) is one that defines the specific rules for the challenge that the company or research center wants to publish. In particular it is the document where you set the conditions of Intellectual and Industrial Challenge. Please read them carefully.

The Challenge Specific Agreement will be adapted to the type of challenge chosen by the solver (or pre-competitive ideas). There is also the possibility that the company or research center incorporates its own Challenge Specific Agreement. Depending on the type of challenge and in case the company or research center did not include other specific conditions, the general rules of intellectual property of the Challenge will be as follows:

  • Challenge ideas: the company or research center will be granted a royalty-free, perpetual, nonexclusive license to use any information of the solution.
  • Desafíos precompetitivos: El desafío requiere la concesión de sus derechos exclusivos de propiedad intelectual e industrial (PI). Si los derechos de propiedad intelectual no pueden ser transferidos por el solucionador en exclusiva, la empresa podría considerar un acuerdo de licencia para un premio parcial.

Precompetitive Challenges: The challenge requires granting of exclusive intellectual property rights (IP). If intellectual property rights can not be transferred by the Solver exclusively, the company or research center might consider a licensing agreement for a partial award.

9. Acceptance

Challenges specific agreements that are proposed by the company or research center will be reviewed by e-OpenFunding Management Services Innnovación, SL to check compliance with the Terms of Use of this Web site.