Differentiation Culture of Hematopoietic Stem Cells from Embryonic Stem Cells.
- Author:
Suk Jin KIM
1
;
Byung Soo KIM
;
Suck Won RYU
;
Hwa Jung SUNG
;
Kyung Hwa PARK
;
In Keun CHOI
;
Sang Chul OH
;
Jae Hong SEO
;
Chel Won CHOI
;
Sang Won SHIN
;
Yeul Hong KIM
;
Jun Suk KIM
Author Information
1. Department of Internal Medicinel, Korea University College of Medicine, Seoul, Korea. kbs0309@korea.ac.kr
- Publication Type:Original Article
- Keywords:
Embryonic stem cell;
Embryonic body;
Hematopoietic stem cell
- MeSH:
Embryonic Stem Cells*;
Hematopoietic Stem Cells*;
Humans
- From:Korean Journal of Hematology
2004;39(2):78-85
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: This 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)x10(4), 1.8 (1.4-2.6)x10(4), 0.6 (0.5-0.7)x10(4) and the median count of CFUs was 0.5 (0.2-0.8)x10(2), 1.0 (0.6-1.3)x10(2), 0.2(0.1-0.4)x10(2) 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 stem cells is between 3 to 7 days of LTC-IC assay.
