Optimizing in vitro conditions for immunomodulation and expansion of mesenchymal stromal cells
| Autor: | K. Le Blanc, H. Samuelsson, Helena Lönnies, O Ringdén |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Cancer Research
Time Factors Stromal cell Cell Survival Lymphocyte Immunology Cell Culture Techniques Lymphocyte proliferation Biology Peripheral blood mononuclear cell Bone Marrow In vivo Freezing medicine Humans Immunology and Allergy Viability assay Phytohemagglutinins Cells Cultured Genetics (clinical) Cell Proliferation Immunosuppression Therapy Transplantation Mesenchymal stem cell Mesenchymal Stem Cells Cell Biology medicine.anatomical_structure Oncology Histocompatibility Leukocytes Mononuclear Cancer research Bone marrow Lymphocyte Culture Test Mixed Stromal Cells |
| Zdroj: | Cytotherapy. 11:129-136 |
| ISSN: | 1465-3249 |
| Popis: | Mesenchymal stromal cells (MSC) can be expanded in vitro for clinical use and have been evaluated in clinical trials as immunosuppressants and to heal damaged tissues. We investigated the impact of freezing and prolonged storage on cell viability, proliferation and immunosuppression in vitro.MSC were expanded from bone marrow (BM) of healthy subjects according to standard protocols in the presence of fetal calf serum (FCS). The immunosuppressive potential of MSC was analyzed in mixed lymphocyte cultures (MLC) and after stimulation with phytohemagglutinin (PHA).Expansion of frozen mononuclear cells (MNC) diminished MSC yield after expansion compared with plating of fresh MNC. MSC derived from frozen MNC were also less immunosuppressive. MSC harvested at various passages after expansion in vivo suppressed lymphocyte proliferation equally. Pooling of MSC from several donors generated higher and more stable suppression in both MLC and after PHA stimulation. After passage 1, plating at lower densities in 20% FCS increased cell expansion of MSC up to 25-fold compared with standard conditions.MNC should not be frozen prior to MSC expansion. Decreased replating density and increased FCS levels generate higher numbers of MSC. Freezing of ex vivo-expanded MSC for30 months did not affect cell viability or the ability to suppress lymphocyte proliferation. For effective immunosuppression in vitro MSC should be stored for less than 6 months and pooled from two or three donors. |
| Databáze: | OpenAIRE |
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