The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). The use of the human umbilical cord tissue from a healthy donor who gave birth and signed informed consent in Renmin Hospital was supported by the Institutional Ethics Review Board of Renmin Hospital of Wuhan University (Permit Number: WDRY2019-G001). UC-MSCs were isolated, cultured and identified as described in our previous study (19 (link)). Briefly, the umbilical cord was cut into small pieces and cultured in serum-free medium (UltraCULTURE, Walkersville Lonza, MD) supplemented with serum alternatives (Life Science, France) and L-Glutamine at 37 °C in a humidified atmosphere containing 5% CO2. The fifth generations of cells were used in this animal experiment. UC-MSCs were characterized for surface marker expression and differentiation potential of adipogenesis, osteogenesis, and chondrogenesis as described previously (19 (link)). For the production of UC-MSCs condition medium (UC-MSCs-CM), the supernatant of UC-MSCs was collected and concentrated 25-fold using ultrafilter (Millipore, Bedford, MA), when the confluence reached about 80%.
Protocol detail
Isolation and Characterization of UC-MSCs
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Adipogenesis
Atmosphere
Birth
Chondrogenesis
Condition medium
Cultured medium
Donor
Human
L glutamine
Osteogenesis
Serum
Tissue
Umbilical cord
Corresponding Organization : Wuhan University
Other organizations : Wuhan Institute of Bioengineering, Wuhan Donghu University
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Variable analysis
independent variables
- Isolation and culture of UC-MSCs
- Production of UC-MSCs condition medium (UC-MSCs-CM)
- Surface marker expression of UC-MSCs
- Differentiation potential of UC-MSCs (adipogenesis, osteogenesis, chondrogenesis)
- Serum-free medium (UltraCULTURE, Walkersville Lonza, MD) supplemented with serum alternatives (Life Science, France) and L-Glutamine
- Incubation at 37 °C in a humidified atmosphere containing 5% CO2
- Use of fifth generation of UC-MSCs
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