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Updated: Sep 16, 2025
Isolation of Human BAMBIhighMFGE8high Umbilical Cord-Derived Mesenchymal Stromal Cells
Published on: January 10, 2025
Comparison between isolation protocols highlights intrinsic variability of human umbilical cord mesenchymal cells
Fernanda Vieira Paladino1, Joana Silveira Peixoto-Cruz1,2, Carolina Santacruz-Perez1
1Hospital Israelita Albert Einstein, IIEP, São Paulo, Brazil.
Abstract:
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.
Insights
Mesenchymal stem cells (MSCs) from umbilical cord wall have limited lifespan in vitro. Enzymatic isolation protocol II yielded MSCs with superior proliferation and extended culture lifespan, crucial for therapeutic applications.
Area of Science:
- Cell Biology
- Stem Cell Research
- Tissue Engineering
Background:
- Mesenchymal stem cells (MSCs) are multipotent cells with therapeutic potential but have limited in vitro lifespan.
- Human umbilical cord wall MSCs are easily accessible and proliferate rapidly, but their expansion and aging characteristics are not fully understood.
- Variability in individual MSC samples can impact their therapeutic efficacy.
Purpose of the Study:
- To compare different isolation protocols for human umbilical cord wall MSCs.
- To identify the protocol yielding the highest number of viable cells with optimal proliferation capacity.
- To evaluate the impact of isolation methods on MSC expansion, senescence, and therapeutic potential.
Main Methods:
- Three isolation protocols were compared: two enzymatic and one explant method.
- Isolated MSCs were assessed for proliferation rate, differentiation potential, and cell surface marker phenotype.
- Cellular behavior, including population doubling times and senescence, was monitored across passages.
Main Results:
- All isolation methods yielded MSCs with standard phenotypic and differentiation characteristics after passage 2.
- Enzymatic isolation protocol II resulted in MSCs that reached later passages and had shorter doubling times, indicating a longer culture lifespan.
- Explant-derived cells senesced earlier than those from enzymatic protocols, regardless of initial proliferation rates.
- Significant variability in proliferation rates and senescence points was observed among individual MSC samples, irrespective of the isolation protocol.
Conclusions:
- Isolating MSCs directly from Wharton's jelly using enzymatic protocol II is advantageous for maximizing cell yield and extending in vitro lifespan.
- Individual umbilical cord samples exhibit unique expansion and aging behaviors, necessitating personalized approaches for therapeutic applications.
- Understanding sample-specific variability is critical for successful clinical translation of MSC-based therapies.
