Use this URL to cite or link to this record in EThOS:
Title: Differentiation of murine embryonic cells towards the haematopoietic cell lineage using the HepG2 conditioned medium and encapsulation in a rotating wall vessel bioreactor
Author: Fauzi, Iliana
ISNI:       0000 0004 2712 4433
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2011
Availability of Full Text:
Access from EThOS:
Access from Institution:
Embryonic stem cells (ESCs) are known for their unique property to be maintained almost indefinitely in an undifferentiated, proliferating state with the potential to give rise to all types of cells. Current established protocols for the culture and differentiation of ESCs are cumbersome and inefficient involving three stages: a) maintenance or expansion of undifferentiated ESCs, b) spontaneous differentiation through formation of embryoid bodies (EBs), and c) dissociation of the EBs and replating leading to the terminal differentiation to the desired lineages. One of the major challenges in the use of ESCs for cell therapy is controlling their differentiation pathways. Optimal culture conditions and requirement as well as precise differentiation mechanisms and cellular interactions within EBs are still not well characterised resulting in sub-optimal control of homogenous differentiation especially due to the formation of all three germ layers. Attempts on developing an efficient culture protocol have been widely reported in order to overcome the limitations. Recent research approaches have shown that treatment with conditioned medium derived from HepG2, a human hepatocarcinoma cell line enhances the formation of multipotent mesodermal progenitors from ESCs. This promotes a greater control of ESC differentiation in a lineage-specific fashion possibly resulting in efficient haematopoietic differentiation. In this study, we have developed an integrated, single step bioprocess for ESCs hematopoietic differentiation that: a) uses HepG2-conditioned medium (HepG2-CM), that stimulates mesoderm formation, b) facilitates three dimensional (3D) culture through encapsulation of undifferentiated ESCs in hydrogels, c) bypasses EB formation, and d) involves culture in a rotating wall vessel bioreactor that does not require passaging of the cells and is scalable and automatable. In conclusion, this thesis reports the development of a novel culture system for the efficient single-step haematopoietic differentiation of ESC resulting in a reproducible, scalable, high-intensity culture system of mESCs for ex-vivo blood manufacture.
Supervisor: Mantalaris, Athanasios ; Panoskaltsis, Nicki Sponsor: Ministry of Higher Education Malaysia ; Richard Thomas Leukaemia Fund ; Northwick Park Hospital Leukaemia Research Trust Fund
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral