Dr Susan Jehangir1, Dr Seng Chua2, Dr John Harvey1, Dr Peter Maitz3, Dr Zhe Li3
1Children’s Hospital At Westmead, Westmead, Australia, 2Department of Women’s and new born health Westmead Hospital, Westmead, Australia, 3Burns Unit, Concord Hospital, , Concord, Australia
There is a renewed interest in the use of Amniotic membrane (AM) as a scaffold for the ex vivo expansion and delivery of cells. This study investigates the issues with harvesting and effects of different preservation procedures on the properties of the AM to serve as a scaffold for the adherence and expansion of stem cells.
Fresh AM was harvested from seronegative term pregnant mothers undergoing elective Caesarean section deliveries under sterile conditions. Decellularization of human amnion HAM was carried out by incubating the tissue in hypotonic tris-EDTA buffer containing EDTA and aprotinin at 4 oC for 16 h. Then tissue was then transferred to a 0.03% SDS solution and incubated, with shaking, for 24 h at room temperature. After treatment with SDS, the tissue was washed thoroughly in buffered saline and either cryopreserved or lyophilised for storage. Cells were seeded onto the epithelial side with medium changed every 2 days. The effects of cryopreservation and lyophilisation were evaluated on mechanical and histological characteristics of the AM and its effects on cell adherence and expansion.
Preliminary results favour the freeze dried AM for cell adhesion and expansion. Final results awaited
Visiting from Christian Medical College, Vellore as a short term trainee. Fellow at the Children’s Hospital at Westmead with a special interest in the use of amniotic membrane in burns