TY - JOUR
T1 - Reversible switching of expression of c-kit and Pax-5 in immature hematopoietic progenitor cells by stromal cells
AU - Okubo, Tadashi
AU - Yanai, Nobuaki
AU - Ikawa, Shuntaro
AU - Obinata, Masuo
N1 - Funding Information:
This work was supported by a Grant-in-Aid from the Ministry of Education, Science, Sports, Culture and Technology of Japan.
PY - 2002/10/1
Y1 - 2002/10/1
N2 - Objective. Bone marrow stromal cells provide the microenvironment for self-renewal and differentiation of hematopoietic stem/progenitor cells through complex cell-cell interaction. To elucidate the regulatory mechanisms of hematopoiesis by stromal cells, we established a novel stroma-dependent hematopoietic cell line and explored the phenotypic changes regulated by the two stromal cells. Materials and Methods. DFC-28 cells clonally established from long-term bone marrow culture of C57BL/6 mice were sustained by coculture on MSS62 cells (mouse spleen stromal cell line). When DFC-28 cells were transferred to TBR31-1 cells (mouse bone marrow stromal cell line), their phenotypic changes were analyzed by flow cytometry and reverse transcriptase polymerase chain reaction. Results. DFC-28 cells on MSS62 cells exhibited surface phenotypes of the immature hematopoietic progenitor cells (Lin-AA4.1+c-kit+Sca-1-). By stroma-replacement from MSS62 cells to TBR31-1 cells, DFC-28 cells were differentiated into very early B-lymphoid stage characterized by c-kit down-regulation and induction of BP-1 and B-lymphoid-associated genes (Pax-5, CD19, TdT, Rag-1, and Rag-2). In addition, the differentiation phenotypes reverted to the immature state characterized by c-kit induction and down-regulation of BP-1 and B-lymphoid-associated genes by replacing stroma back to MSS62 from TBR31-1. Interleukin-7 stimulation and conditioned medium of TBR31-1 cells were ineffective in converting the differentiation phenotypes of DFC-28 cells. Conclusion. The results demonstrate that the differentiation phenotypes and growth potential of stroma-dependent hematopoietic progenitor cells we established could be reversibly controlled via direct contact with stromal cells in the microenvironment.
AB - Objective. Bone marrow stromal cells provide the microenvironment for self-renewal and differentiation of hematopoietic stem/progenitor cells through complex cell-cell interaction. To elucidate the regulatory mechanisms of hematopoiesis by stromal cells, we established a novel stroma-dependent hematopoietic cell line and explored the phenotypic changes regulated by the two stromal cells. Materials and Methods. DFC-28 cells clonally established from long-term bone marrow culture of C57BL/6 mice were sustained by coculture on MSS62 cells (mouse spleen stromal cell line). When DFC-28 cells were transferred to TBR31-1 cells (mouse bone marrow stromal cell line), their phenotypic changes were analyzed by flow cytometry and reverse transcriptase polymerase chain reaction. Results. DFC-28 cells on MSS62 cells exhibited surface phenotypes of the immature hematopoietic progenitor cells (Lin-AA4.1+c-kit+Sca-1-). By stroma-replacement from MSS62 cells to TBR31-1 cells, DFC-28 cells were differentiated into very early B-lymphoid stage characterized by c-kit down-regulation and induction of BP-1 and B-lymphoid-associated genes (Pax-5, CD19, TdT, Rag-1, and Rag-2). In addition, the differentiation phenotypes reverted to the immature state characterized by c-kit induction and down-regulation of BP-1 and B-lymphoid-associated genes by replacing stroma back to MSS62 from TBR31-1. Interleukin-7 stimulation and conditioned medium of TBR31-1 cells were ineffective in converting the differentiation phenotypes of DFC-28 cells. Conclusion. The results demonstrate that the differentiation phenotypes and growth potential of stroma-dependent hematopoietic progenitor cells we established could be reversibly controlled via direct contact with stromal cells in the microenvironment.
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U2 - 10.1016/S0301-472X(02)00899-8
DO - 10.1016/S0301-472X(02)00899-8
M3 - Article
C2 - 12384151
AN - SCOPUS:0036792029
VL - 30
SP - 1193
EP - 1201
JO - Experimental Hematology
JF - Experimental Hematology
SN - 0301-472X
IS - 10
ER -