Epidermal stem cells are preserved during commercial-scale manufacture of a bilayered, living cellular construct (Apligraf®)
| Autor: | Mark W. Carlson, Katie Faria, Jonathan Leman, Yulia Shamis, Jonathan A. Garlick, Vincent Ronfard |
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| Rok vydání: | 2010 |
| Předmět: |
Cell Survival
Cell Population Biomedical Engineering Mice Nude Bioengineering Biology Biochemistry Biomaterials Mice Retrovirus Nude mouse In vivo Materials Testing medicine Animals Humans education Skin Artificial education.field_of_study Stem Cells biology.organism_classification In vitro Cell biology Transplantation medicine.anatomical_structure Epidermal Cells Immunology Collagen Stem cell Epidermis |
| Zdroj: | Tissue engineering. Part A. 17(3-4) |
| ISSN: | 1937-335X |
| Popis: | It is unknown if epidermal stem cells are maintained during the commercial-scale manufacture of Apligraf, a bilayered living cellular construct (BLCC). To answer this question, we genetically marked replicating keratinocytes, derived from production-scale expansion of working cell banks, in two-dimensional culture with a beta-galactosidase-expressing retrovirus and monitored their fate after incorporation into BLCC and subsequent in vivo transplantation to a nude mouse. Histological analysis of BLCCs showed distinct beta-galactosidase-positive clusters similar to clonal proliferation units visible 8-32 weeks after grafting. Keratinocytes recovered from grafts at week 32 were expanded in vitro in two-dimensional culture, and clonal growth of recovered cells was then compared to the original pregraft population of keratinocytes by colony-forming efficiency (CFE) assays. The CFE of the cells regrown from the grafts was similar to pregraft CFEs (45% and 40%, respectively). Cells regrown from the grafts were then used to produce a second BLCC and generated a well-differentiated epithelium that was histologically similar to pregraft BLCC. These findings provide clear evidence that epidermal stem cells were sustained during the process of large-scale tissue fabrication and that the process of isolation and expansion of cells in BLCC construction retains viable stem cells. |
| Databáze: | OpenAIRE |
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