Affect of hypoxic conditions on

mesenchymal stem cell expansion

and chondrogenic differentiation
NIKITA MILANI
UNIVERSITY OF WASHINGTON
DR. YASUYUKI SAKAI
INSTITUTE OF INDUSTRIAL SCIENCE

Background
Cartilage tissue degrades over time
Stem cell therapies are emerging as treatment

MSCs acquired from hospital waste (polydactyly)

thighbone

Aging
Overwork

shinbone
Sharp Pain Joint deformity

Project
Problem: MSCs lose potency and viability

over time
Determine affect of hypoxic conditions and substrate
material on MSC expansion and differentiation

5%, 10%, 20% oxygen

PDMS, TCPS

24 well PDMS plate (left) and 10 cm TCPS dish (right)

Results: Expansion
TCPS

PDMS
Cell counting

20%

10%

5%
Figure 1: Images of MSCs in
expansion phase taken on
day 3

25000
20000
15000
Cell density (cells/cm^2) 10000
5000
0

TCPS
PDMS

Oxygen concentration
Figure 2: Graph comparing the number
of cells cultured in different conditions
on day 3

Results: Differentiation

Conclusion
Low oxygen improved cell growth rate and

differentiation potential
PDMS induced chrondrogenic metabolism earlier in
differentiation
PDMS at 5% oxygen best preserved MSC

potency and viability

Thank you for your attention!

References
Pattappa, G., HK Heywood, JD De Bruijn, and DA Lee. "The

Metabolism of Human Mesenchymal Stem Cells during

Proliferation and Differentiation." J Cell Physiol(2011)
Sato, M., M. Yamato, K. Hamahashi, T. Okano, and J.
Mochida. "Articular Cartilage Regeneration Using Cell Sheet
Technology." The Anatomical Record (2014)
Drela, K., A. Sarnowska, P. Siedlecka, I. SzablowskaGadomska, B. Lukomska, and K. Domanska-Janik. "Low
Oxygen Atmosphere Facilitates Proliferation and Maintains
Undifferentiated State of Umbilical Cordmesenchymal Stem
Cells in an Hypoxia Inducible Factor-dependent
Manner." Cytotherapy (2014)