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Updated: Jul 3, 2026
Recombinant Collagen I Peptide Microcarriers for Cell Expansion and Their Potential Use As Cell Delivery System in a Bioreactor Model
Published on: February 7, 2018
The extended serial subculture of human diploid fibroblasts on microcarriers using a new medium supplement
S P Forestell1, N Kalogerakis, L A Behie
1The Pharmaceutical Production Research Facility (PPRF), Faculty of Engineering, The University of Clagary, Calgary, Alberta, Canada.
Abstract:
Human diploid fibroblasts serially passaged on microcarriers exhibit a decrease in their proliferative capacity with each transfer from microcarrier-to-microcarrier. This phenomenon, which does not occur in the same time scale with cells cultured in T-flasks, has been a serious barrier to the systematic utilization of microcarriers in the scale-up of anchorage-dependent human diploid cell cultures. This decreases in cell growth with each passage is shown to be related to the serum content of the medium, with high serum concentrations resulting in a more rapid decrease in cell growth with each serial transfer. As a result, methods for reducing the serum requirement of the cells were investigated. A new medium supplement mixture, PPRF92, has been developed, which allows the serial passaging of MRC5 cells on Cytodex 1 microcarriers through as many as 13 microcarrier-to-microcarrier transfers, and at a serum levels as low as 1%, with no decrease in the proliferative capacity of the cells until they approach their reported population doubling limit. This new supplement mixture is a significant improvement to microcarrier technology in that it enables the use of microcarriers in the early stages of inoculum build-up for the production purposes.
Insights
A new supplement, PPRF92, overcomes microcarrier limitations for human diploid fibroblasts. This breakthrough enables sustained cell proliferation during serial transfers, crucial for cell culture scale-up.
Area of Science:
- Cell Biology
- Biotechnology
- Bioprocess Engineering
Background:
- Serial passaging of human diploid fibroblasts on microcarriers reduces proliferative capacity.
- This phenomenon hinders microcarrier use in scaling up anchorage-dependent cell cultures.
- Reduced cell growth correlates with higher serum concentrations in culture media.
Purpose of the Study:
- Investigate methods to reduce serum requirements for microcarrier-based cell cultures.
- Develop a novel supplement to maintain fibroblast proliferative capacity during serial microcarrier transfers.
- Enhance microcarrier technology for efficient inoculum build-up in cell production.
Main Methods:
- Developed and tested a new medium supplement mixture, PPRF92.
- Utilized Cytodex 1 microcarriers for serial passaging of MRC5 cells.
- Assessed cell proliferative capacity across multiple microcarrier-to-microcarrier transfers at low serum levels (1%).
Main Results:
- PPRF92 enabled up to 13 serial microcarrier transfers of MRC5 cells.
- Sustained proliferative capacity was observed even at 1% serum concentration.
- No decrease in cell growth occurred until cells approached their population doubling limit.
Conclusions:
- PPRF92 significantly improves microcarrier technology for cell culture scale-up.
- The supplement overcomes previous limitations related to reduced cell proliferation.
- Enables efficient early-stage inoculum build-up for bioproduction using microcarriers.
