Enhanced isolation of fibroblasts from human skin explants
| Autor: | Christine E. Napier, Roger R. Reddel, Antony W. Braithwaite, Amy Y.M. Au, Jane R. Noble, Lily I. Huschtscha, Hamish Campbell, Kylie Bower |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Adolescent Population Cell Cell Culture Techniques Human skin Cell Separation Biology General Biochemistry Genetics and Molecular Biology Cryopreservation Andrology Mice Young Adult medicine Animals Humans education Aged Skin education.field_of_study integumentary system Lentivirus Fibroblasts Middle Aged Telomere medicine.anatomical_structure Cell culture Cryopreserved Tissue Female Biotechnology Explant culture |
| Zdroj: | BioTechniques. 53(4) |
| ISSN: | 1940-9818 |
| Popis: | Here we describe a method for growing fibroblasts from human skin explants that increases the number of cells obtained by up to two orders of magnitude, thus increasing the amount of material available for research and diagnostic purposes and potentially for cell-based therapies. Explants can be transferred sequentially up to 80 times, if required, at which point the explants appear to be completely depleted of fibroblasts. Utilizing skin samples obtained from 16 donors, aged 18-66 years old, the first 20 transfers produced cultures with lifespan and growth characteristics that were all very similar to each other, but the cultures derived from later transfers had a decreasing replicative capacity. Final cumulative population doublings did not correlate with donor age, but correlated positively with the telomere length at early passage. We also demonstrated that explants can be transduced directly by lentiviral infection, and that cryopreserved tissue can be explanted successfully using this procedure. |
| Databáze: | OpenAIRE |
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