Expansion of mesenchymal stem cells on fibrinogen-rich protein surfaces derived from blood plasma
John D Kisiday1, Benjamin W Hale, Jorge L Almodovar
1Department of Clinical Science, Colorado State University, Fort Collins, CO, USA. john.kisiday@colostate.edu
Abstract:
Mesenchymal stem cells (MSCs) are present in low density in bone marrow and culture expansion is necessary to obtain sufficient numbers for many proposed therapies. Researchers have characterized MSC growth on tissue culture plastic (TCP), although few studies have explored proliferation on other growth substrates. Using adult equine MSCs, we evaluated proliferation on fibrinogen-rich precipitate (FRP) surfaces created from blood plasma. When seeded at 1 × 10(4) cells/cm(2) and passaged five times over 10 days, MSCs on FRP in medium containing fibroblast growth factor 2 (FGF2) resulted in a ∼2.5-fold increase in cell yield relative to TCP. In FGF2-free medium, FRP stimulated a 10.4-fold increase in cell yield over TCP after 10 days, although control cultures maintained in FGF2 on TCP demonstrated that the stimulatory effect of FRP was not as lasting as that of FGF2. Chondrogenic cultures demonstrated that FRP did not affect differentiation. On TCP, MSCs seeded at 500 cells/cm(2) experienced a 4.6-fold increase in cell yield over cultures seeded at 1 × 10(4) cells/cm(2) following 10 days of expansion. In 500 cells/cm(2) cultures, FRP stimulating a two-fold increase in cell yield over TCP without affecting differentiation. Low-density FRP cultures showed a more even distribution of cells than TCP, suggesting that FRP may accelerate proliferation by reducing contact inhibition that slows proliferation. In addition, FRP appears capable of binding FGF2, as FRP surfaces pre-conditioned with FGF2 supported greater proliferation than FGF2-free cultures. Taken together, these factors indicate that substrates obtained from simple and inexpensive processing of blood enhance MSC proliferation and promote efficient coverage of expansion surfaces.
Insights
Fibrinogen-rich precipitate (FRP) from blood plasma enhances mesenchymal stem cell (MSC) proliferation on culture surfaces. This inexpensive substrate increases cell yield and coverage, potentially reducing contact inhibition and binding growth factors.
Area of Science:
- Cell Biology
- Biomaterials Science
- Regenerative Medicine
Background:
- Mesenchymal stem cells (MSCs) require expansion for therapeutic use.
- Tissue culture plastic (TCP) is the standard substrate, but alternatives are underexplored.
- Equine MSCs were used to investigate novel growth substrates.
Purpose of the Study:
- To evaluate the proliferation of equine MSCs on fibrinogen-rich precipitate (FRP) surfaces.
- To compare MSC expansion on FRP versus traditional TCP.
- To assess the impact of FRP on MSC differentiation and cell distribution.
Main Methods:
- Adult equine MSCs were cultured on FRP and TCP at different seeding densities.
- Cell proliferation was measured over 10 days with and without fibroblast growth factor 2 (FGF2).
- Chondrogenic differentiation potential was assessed for MSCs cultured on FRP.
Main Results:
- FRP significantly increased MSC yield compared to TCP, especially in FGF2-free medium (10.4-fold increase).
- FRP promoted more even cell distribution, suggesting reduced contact inhibition.
- FRP did not negatively affect MSC chondrogenic differentiation and could bind FGF2, enhancing proliferation.
Conclusions:
- FRP is a cost-effective blood-derived substrate that enhances MSC proliferation.
- FRP offers advantages over TCP for MSC expansion, including improved cell coverage and potential for growth factor binding.
- FRP shows promise as a biomaterial for improving MSC-based therapies.
