title: Translation of cell therapy into clinical practice: validation of an application procedure for bone marrow progenitor cells and platelet rich plasma.
abstract: OBJECTIVE
  Tissue regeneration can be improved by local application of autologous bone marrow derived progenitor cells (BMSC) and platelet rich plasma (PRP). However, there is a lack of standardized application procedures for clinical use. Therefore, a technique in accordance with the guidelines for advanced therapies medical products of the European Medicine Agency was developed and established.
  
  
  METHODS
  In detail, a process for the isolation and formulation of autologous bone marrow cells (BMC) and PRP in a clinical setting was validated. To investigate the influence of storage time and temperature on gel formation and gel stability, different concentrations of BMC were stored with and without additional platelets, thrombin and fibrinogen and analyzed over a period of 28 days. In addition, cell vitality using a live-dead staining and migration ability of human mesenchymal stem cells (hMSC) in the gel clot was investigated.
  
  
  RESULTS
  For an optimized stable gel clot, human BMC and PRP should be combined with 10% to 20% fibrinogen (9 mg/mL to 18 mg/mL) and 5% to 20% thrombin (25 I.E. to 100 I.E.). Both freshly prepared and stored cells for 1 to 7 days had a stable consistence over 28 days at 37°C. Different platelet concentrations did not influence gel clot formation. The ratio of living cells did not decrease significantly over the observation period of 5 days in the live-dead staining.
  
  
  CONCLUSIONS
  The study identified an optimal gel texture for local application of BMC and PRP. Seeded hMSC could migrate therein and were able to survive to initiate a healing cascade.
authors.affiliation: University Clinic Dresden, Orthopaedic and Traumatology Centre (OUC), Dresden - Germany.|Centre for Translational Bone, Joint and Soft Tissue Research, Dresden - Germany.
authors.forename: Joerg
authors.initials: J
authors.lastname: Nowotny
authors.affiliation: Centre for Translational Bone, Joint and Soft Tissue Research, Dresden - Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Farack
authors.affiliation: Centre for Translational Bone, Joint and Soft Tissue Research, Dresden - Germany.
authors.forename: Corina
authors.initials: C
authors.lastname: Vater
authors.affiliation: Red Cross Blood Transfusion Service North-East, Dresden - Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Johnsen
authors.affiliation: Centre for Translational Bone, Joint and Soft Tissue Research, Dresden - Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Gelinsky
authors.affiliation: Red Cross Blood Transfusion Service North-East, Dresden - Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Tonn
authors.affiliation: Orthopaedic Surgery Center (OCC), Tübingen - Germany.
authors.forename: Philip
authors.initials: P
authors.lastname: Kasten
mesh_terms: D064592:Autografts; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D017690:Cell Transplantation / Q000379:methods*; D006801:Humans; D053657:Platelet-Rich Plasma; D013234:Stem Cells; D057170:Translational Research, Biomedical