Comparison between isolation protocols highlights intrinsic variability of human umbilical cord mesenchymal cells

Fernanda Vieira Paladino, Joana Silveira Peixoto-Cruz, Carolina Santacruz-Perez, Anna Carla Goldberg

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)


Mesenchymal stem cells (MSCs) though multipotent exhibit limited lifespan in vitro, with progressive reduction in capacity for self-renewal leading to irreversible arrest of cell division, which limits their use for therapeutic purposes. Human umbilical cord wall MSCs are easy to process and proliferate rapidly in culture, but variability of individual samples and impact upon in vitro expansion and aging processes is unknown. We compared isolation protocols to determine which one yields the highest number of viable cells with the best proliferation capacity. Three different protocols were tested: two were enzymatic procedures and one explant method. Isolated cells were evaluated in terms of proliferation, differentiation capacity, and phenotype. All samples were processed using one or more protocols. After passage 2 adherent cells displayed standard phenotypic and differentiation characteristics of MSCs, but our results show that isolating cells directly from Wharton’s jelly is more advantageous. Cells obtained from explants presented similar characteristics to those from enzymatic protocols, but always reached proliferation arrest earlier, irrespective of initial population doubling times. From the same sample, cells obtained with enzymatic protocol ii reached later passages while exhibiting shorter doubling times in culture than cells from other protocols, that is, took longer to reach senescence. More important, each individual MSC sample exhibited different population doubling rates and reached senescence at different passages, irrespective of protocol. Thus, even when in strict conformity with procedures and quality control, each cord sample shows a unique behavior, a finding that should be taken into account when planning for therapeutic approaches.

Original languageEnglish
Pages (from-to)123-136
Number of pages14
JournalCell and Tissue Banking
Issue number1
Publication statusPublished - 1 Mar 2016


  • Isolation methods
  • Mesenchymal stem cells
  • Replicative senescence
  • Umbilical cord


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