Development of the experimental advanced-therapy medicinal product: isolation, cultivation and characterization of muscle derived stem cells
| Author | Affiliation |
|---|---|
Mauricas, Mykolas | Valstybinis mokslinių tyrimų institutas Inovatyvios medicinos centras |
Sūdžius, Gintaras | Valstybinis mokslinių tyrimų institutas Inovatyvios medicinos centras |
| Date |
|---|
2014-11-22 |
Bibliogr.: p. 19
Introduction. Adult stem cells hold great potential for regenerative medicine and have significant advantages over embryonic and fetal stem cells.1 One of the main advantages is a possibility to use adult stem cells for autologous therapy, as it eliminates the risk of immune reactions or serious side effects.2 Adult stem cells have been isolated from various tissues.3-6 Muscle-derived stem cells (MDSC) have several advantages over adult stem cell populations, derived from other tissues. MDSCs are multipotent, relatively easy to isolate, biopsy is less traumatic to the patient comparing with the bone marrow biopsy, a small amount of skeletal muscle is needed to isolate a sufficient number of stem cells for treatment purposes.2 Therefore, MDSCs can be used as a potential source for the development of experimental advanced-therapy medicinal products, for various therapeutic indications. Objective. The main objective of this study was to isolate, cultivate and characterize rat MDSCs as an experimental advanced-therapy medicinal product. Methods. Isolation and cultivation of MDSC were performed according to Lavasani M et al. previously published paper1 with some modifications. 2-3 week old Wistar male rat was used for the MDSC isolation. After euthanasia and muscle biopsy, the muscle was washed, dissected from the connective tissue, mechanically crushed and enzymatically digested using collagenase, dispase and trypsin. Cells were platted on collagen type I-coated T-25 flask and incubated at 37°C in a humidified, 5% CO2 incubator for 2 h (labeled pp1). Media with nonadherent cells was transferred into a second T-25 flask, labeled pp2 and incubated for 18 h. On the next day media with nonadherent cells was centrifuged at 930×g at 4°C for 5 min. Supernatant was aspirated, cell pellet was resuspended in 5 ml of media, transferred into the third T-25 flask labeled pp3 and returned to the incubator for 24 h. This pr... [...].