Hematopoietic differentiation of embryoid bodies derived from the human embryonic stem cell line SNUhES3 in co-culture with human bone marrow stromal cells.
Human embryonic stem (ES) cells can be induced to differentiate into hematopoietic precursor cells via two methods: the formation of embryoid bodies (EBs) and co-culture with mouse bone marrow (BM) stromal cells. In this study, the above two methods have been combined by co-culture of human ES-cell-derived EBs with human BM stromal cells. The efficacy of this method was compared with that using EB formation alone. The undifferentiated human ES cell line SNUhES3 was allowed to form EBs for two days, then EBs were induced to differentiate in the presence of a different serum concentration (EB and EB/high FBS group), or co- cultured with human BM stromal cells (EB/BM co-culture group). Flow cytometry and hematopoietic colony-forming assays were used to assess hematopoietic differentiation in the three groups. While no significant increase of CD34+/CD45- or CD34+/CD38- cells was noted in the three groups on days 3 and 5, the percentage of CD34+/CD45- cells and CD34+/ CD38- cells was significantly higher in the EB/BM co-culture group than in the EB and EB/high FBS groups on day 10. The number of colony-forming cells (CFCs) was increased in the EB/BM co-culture group on days 7 and 10, implying a possible role for human BM stromal cells in supporting hematopoietic differentiation from human ES cell-derived EBs. These results demonstrate that co-culture of human ES-cell-derived EBs with human BM stromal cells might lead to more efficient hematopoietic differentiation from human ES cells cultured alone. Further study is warranted to evaluate the underlying mechanism.
Kim
Seok Jin
SJ
Division of Hematology/Oncology, Department of Internal Medicine, Korea University Medical Center, 126-1 Anam-dong 5ga, Sungbuk-gu, Seoul 136-705, Korea.
Kim
Byung Soo
BS
Ryu
Suck Won
SW
Yoo
Ji Hyun
JH
Oh
Jee Hyun
JH
Song
Chang Hee
CH
Kim
Sun Haeng
SH
Choi
Dong Seop
DS
Seo
Jae Hong
JH
Choi
Chul Won
CW
Shin
Sang Won
SW
Kim
Yeul Hong
YH
Kim
Jun Suk
JS
D051997:ADP-ribosyl Cyclase 1 / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D004622:Embryo, Mammalian / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D013234:Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000502:physiology
D018952:Antigens, CD34; D017493:Leukocyte Common Antigens; D051997:ADP-ribosyl Cyclase 1