Culture system for extensive production of CD19+IgM+ cells by human cord blood CD34+ progenitors.
We established a co-culture system with a monolayer of the murine bone marrow (BM) stroma cell line, MS-5, in which human cord blood CD34+ cells differentiated to CD19+ cells. The addition of stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) highly enhanced the production of CD19+ cells. The expansion of the cell numbers was over 10(3)-fold. Furthermore, a significant proportion (<45%) of the cells expressed surface IgM (sIgM) after 5 weeks of co-culture. CD34+CD19- cells also showed a similar development of CD19+ cells and CD19+sigM+ cells. Filter separation of MS-5 cells and CD34+ cells did not inhibit the growth of CD19+ cells. However, when further purified CD34+CD19-CD13- CD33- cells were cultured in the presence of MS-5 cells with or without a separation filter, CD19+ cells did not appear in the non-contact setting. This result suggested that the highly purified CD34+CD19-CD13-CD33- progenitors require the cell-cell contact for the development of CD19+ cells, whereas other CD34+ fractions contain progenitors that do not require the contact. This co-culture system should be useful for the study of early human B-lymphopoiesis.
Ohkawara
J I
JI
R&D Center, TERUMO Corporation, Nakai, Kanagawa, Japan.
Ikebuchi
K
K
Fujihara
M
M
Sato
N
N
Hirayama
F
F
Yamaguchi
M
M
Mori
K J
KJ
Sekiguchi
S
S
D000818:Animals; D018941:Antigens, CD19 / Q000276:immunology*; D018952:Antigens, CD34 / Q000097:blood* / Q000502:physiology; D001402:B-Lymphocytes / Q000166:cytology; D001709:Biotechnology / Q000379:methods; D002450:Cell Communication / Q000502:physiology; D002460:Cell Line; D018920:Coculture Techniques; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007075:Immunoglobulin M / Q000096:biosynthesis* / Q000097:blood*; D051379:Mice; D008809:Mice, Inbred C3H; D019089:Stem Cell Factor / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology
D018941:Antigens, CD19; D018952:Antigens, CD34; D007075:Immunoglobulin M; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor