BACKGROUND:
his study was designed to verify the effective culture condition for the differentiation of human hematopoietic stem cells from SNU-3 embryonic stem cell line.
METHODS :
Control group was that of which embryonic stem cells were directly cultured to LTC-IC assay. Study group was that of which an embryonic body manufactured from embryonic stem cells was cultured to LTC-IC assay. CD34＋ cells were separated by MACS method at 1, 3, 5, 7, 10, 14, 21 days of LTC-IC assay in both groups. Thereafter, CD34＋ cell were cultured to semisolid methyl-cellulose media to count CFUs. CD34＋CD45- cell percentage was measured with FACS method at 5 days of LTC-IC assay. This study was repeated for 14 times.
RESULTS :
In control group, CD34＋ cells were hardly separated in any period of LTC-IC assay. In study group, the median count of CD34＋ cells was 0.7 (0.4-1.2)×104, 1.8 (1.4- 2.6)×104, 0.6 (0.5-0.7)×104 and the median count of CFUs was 0.5 (0.2-0.8)×102, 1.0 (0.6-1.3)×102, 0.2(0.1-0.4)×102 at 3, 5, 7 days of LTC-IC assay, respectively. Median CFUs count per CD34＋ cell was 0.0071, 0.0056, 0.0033 at 3, 5, 7 days of LTC-IC assay, respectively. In study group, the count of CD34＋ cells and CFUs was significantly higher at 5 days of LTC-IC assay than at any other days (P<0.01). CFUs count per CD34＋ cell was significantly higher at 3 days of LTC-IC assay than at any other days (P<0.01). CD34＋CD45- cells detected by FACS method of study group(1.16%(0.92-1.97) was significantly higher than that of control group (0.09% (0.00-0.23)) (P<0.01).
CONCLUSION :
The differentiation of hematopoietic stem cells from SNU-3 embryonic stem cell line is effective in the condition of which an embryonic body manufactured from embryonic stem cells is cultured to LTC-IC assay. The period of which embryonic stem cells differentiate to hematopoietic stemcells is between 3 to 7 days of LTC-IC assay

Keywords: Embryonic stem cell, Embryonic body, Hematopoietic stem cell