In vitro erythropoiesis of cord blood haematopoietic stem cells (HSCs) to produce
fully enucleated red blood cells could provide an alternate resource for the erythrocyte.
However, haematopoietic processes are highly complex and dynamic; defining process
requirements to produce reproducible cells of high purity and yield is not an easy task. One
major obstacle is the lack of knowledge in process characteristics. Design of experiments
(DOE) is proposed as a tool to unveil process complexities that exists in HSC cultures.
Characterisation and optimisation of in vitro erythropoiesis as a single-step culture is first
performed via a simple DOE experimental strategy. The optimised DOE culture produced
significantly better results (higher growth and faster maturation) than other single-step
cultures. Subsequently, use of DOE to reveal in vitro process dynamics was attempted. This
study was much more challenging and the repeatability of DOE process models was
compromised in some cases. Process control of HSC culture bioprocesses is required for the
delivery of reliable cell culture products suited for clinical applications. The availability of a
convenient and economical online real-time process monitoring system can provide the
means to translate stem cell culture bioprocesses from the bench-side into manufacturing
production. The design and integration of such a system capable of simultaneous process
monitoring of multiple analytes (ammonia, pH and oxygen) is presented. Operational and
functional stability of this unique online real-time in situ monitoring platform was achieved.
Stability of oxygen and ammonia sensors was achieved for up to three and six days
respectively but biocompatibilities of both sensors require some improvements. Sensors of
pH were biocompatible but their stability in cell culture is required.