Preclinical ex vivo expansion of cord blood hematopoietic stem and progenitor cells: duration of culture; the media, serum supplements, and growth factors used; and engraftment in NOD/SCID mice

Abstract

The data supported the strategy of expansion. The optimized condition may be applicable to clinical expansion for the abrogation or reduction of posttransplant cytopenia.

Cobblestone area-forming cells, long-term culture-initiating cells and NOD/SCID repopulating cells in human neonatal blood: a comparison with umbilical cord blood

“…12 In this study, we expanded cryopreserved and thawed NB at a condition optimized for clinical appli- cations, using TPO, SCF, FL and G-CSF in a serum-free medium. 16 Our results demonstrated that NB was significantly expanded to produce early and committed progenitor cells of the myeloid and megakaryocytic lineages. More significantly, SRC were present in the expanded product as revealed by huCD45 + cells in the BM, spleen and PB of the recipients, although in a reduced quantity.…”

“…16,17 NOD/LtSZ-scid/scid mice were purchased from The Walter and Eliza Hall Institute of Medical Research (Victoria, Australia), bred and maintained in the Laboratory Animal Services Centre at the Chinese University of Hong Kong. Animals were handled under sterile conditions and maintained in microisolator cages.…”

“…1 ϫ 10 5 cells were cultured for 14 days in 35 mm plates with 1 ml IMDM containing 10% BSA, 10 Ϫ4 m ␤-mercaptoethanol, 10% FCS, 10% bovine plasma (Sigma), 0.34 mg/ml CaCl 2 , 50 ng/ml TPO and 20 ng/ml IL-3 as described previously. 16 After staining with anti-human CD61-FITC antibody (Dako), CFU-MK was scored under a fluorescence microscope.…”

Hematopoietic stem cells from peripheral blood the perspective of non-mobilized peripheral blood

“…A number of serum-free media have been used over the last few years with different results [59][60][61][62]. For obtaining sufficient numbers of progenitor cells for transplant, FCS [63] and autologous plasma [60,61] have been used in clinical expansion protocols.…”

In-vitro Behavior of Human Umbilical Cord Blood Stem Cells Towards Serum Based Minimal Cytokine Growth Conditions

“…Some of these most frequently used are SCF, Flt3L, Thrombopoietin (TPO), Interleukin-3 (IL-3), and IL-6. Lam et al (2001) [7] reported that use of SCF ? Flt3L ?…”