Abstract
Objective: The role of interleukin (IL)-7 in human B lymphopoiesis is still controversial. We used an in vitro culture system to verify involvement of IL-7 in development of human pro-B cells from hematopoietic stem cells. Materials and Methods: Human CD34+ bone marrow cells were cultured for 4 weeks on MS-5 mouse stromal cells to induce pro-B cells. Expression of IL-7 receptor α or other B-cell differentiation marker genes on cultured human CD34+bone marrow cells was investigated by reverse transcription polymerase chain reaction (RT-PCR). Colony assay of human CD34+ bone marrow cells was also performed to determine the effect of IL-7 on colony-forming ability. Neutralizing antibody or reagent that eliminates the effect of IL-7 was added to the culture system, and the number of pro-B cells induced was estimated by flow cytometry. Results: RT-PCR analysis revealed mRNA expression of IL-7 receptor α as well as B-cell differentiation marker genes in not only CD19+ pro-B cells but also CD19- CD33- cells induced from CD34+ bone marrow cells after cultivation for 4 weeks on MS-5 cells. Addition of anti-mouse IL-7 antibody, anti-human IL-7 receptor α antibody, or JAK3 kinase inhibitor reduced the number of pro-B cells induced, demonstrating that elimination of IL-7 reduces pro-B-cell development. Addition of anti-mouse IL-7 antibody emphasized the colony-forming ability of burst-forming unit erythroid cells. Conclusions: IL-7 produced by MS-5 cells is required for human pro-B-cell development from CD34+bone marrow cells in our culture system, and IL-7 appears to play a certain role in early human B lymphopoiesis.
| Original language | English |
|---|---|
| Pages (from-to) | 1398-1407 |
| Number of pages | 10 |
| Journal | Experimental Hematology |
| Volume | 35 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - 2007 Sep |
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ASJC Scopus subject areas
- Cancer Research
- Cell Biology
- Genetics
- Hematology
- Oncology
- Transplantation
Cite this
Interleukin-7 contributes to human pro-B-cell development in a mouse stromal cell-dependent culture system. / Taguchi, Tomoko; Takenouchi, Hisami; Shiozawa, Yusuke; Matsui, Jun; Kitamura, Noriko; Miyagawa, Yoshitaka; Katagiri, Yoko U.; Takahashi, Takao; Okita, Hajime; Fujimoto, Junichiro; Kiyokawa, Nobutaka.
In: Experimental Hematology, Vol. 35, No. 9, 09.2007, p. 1398-1407.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Interleukin-7 contributes to human pro-B-cell development in a mouse stromal cell-dependent culture system
AU - Taguchi, Tomoko
AU - Takenouchi, Hisami
AU - Shiozawa, Yusuke
AU - Matsui, Jun
AU - Kitamura, Noriko
AU - Miyagawa, Yoshitaka
AU - Katagiri, Yoko U.
AU - Takahashi, Takao
AU - Okita, Hajime
AU - Fujimoto, Junichiro
AU - Kiyokawa, Nobutaka
PY - 2007/9
Y1 - 2007/9
N2 - Objective: The role of interleukin (IL)-7 in human B lymphopoiesis is still controversial. We used an in vitro culture system to verify involvement of IL-7 in development of human pro-B cells from hematopoietic stem cells. Materials and Methods: Human CD34+ bone marrow cells were cultured for 4 weeks on MS-5 mouse stromal cells to induce pro-B cells. Expression of IL-7 receptor α or other B-cell differentiation marker genes on cultured human CD34+bone marrow cells was investigated by reverse transcription polymerase chain reaction (RT-PCR). Colony assay of human CD34+ bone marrow cells was also performed to determine the effect of IL-7 on colony-forming ability. Neutralizing antibody or reagent that eliminates the effect of IL-7 was added to the culture system, and the number of pro-B cells induced was estimated by flow cytometry. Results: RT-PCR analysis revealed mRNA expression of IL-7 receptor α as well as B-cell differentiation marker genes in not only CD19+ pro-B cells but also CD19- CD33- cells induced from CD34+ bone marrow cells after cultivation for 4 weeks on MS-5 cells. Addition of anti-mouse IL-7 antibody, anti-human IL-7 receptor α antibody, or JAK3 kinase inhibitor reduced the number of pro-B cells induced, demonstrating that elimination of IL-7 reduces pro-B-cell development. Addition of anti-mouse IL-7 antibody emphasized the colony-forming ability of burst-forming unit erythroid cells. Conclusions: IL-7 produced by MS-5 cells is required for human pro-B-cell development from CD34+bone marrow cells in our culture system, and IL-7 appears to play a certain role in early human B lymphopoiesis.
AB - Objective: The role of interleukin (IL)-7 in human B lymphopoiesis is still controversial. We used an in vitro culture system to verify involvement of IL-7 in development of human pro-B cells from hematopoietic stem cells. Materials and Methods: Human CD34+ bone marrow cells were cultured for 4 weeks on MS-5 mouse stromal cells to induce pro-B cells. Expression of IL-7 receptor α or other B-cell differentiation marker genes on cultured human CD34+bone marrow cells was investigated by reverse transcription polymerase chain reaction (RT-PCR). Colony assay of human CD34+ bone marrow cells was also performed to determine the effect of IL-7 on colony-forming ability. Neutralizing antibody or reagent that eliminates the effect of IL-7 was added to the culture system, and the number of pro-B cells induced was estimated by flow cytometry. Results: RT-PCR analysis revealed mRNA expression of IL-7 receptor α as well as B-cell differentiation marker genes in not only CD19+ pro-B cells but also CD19- CD33- cells induced from CD34+ bone marrow cells after cultivation for 4 weeks on MS-5 cells. Addition of anti-mouse IL-7 antibody, anti-human IL-7 receptor α antibody, or JAK3 kinase inhibitor reduced the number of pro-B cells induced, demonstrating that elimination of IL-7 reduces pro-B-cell development. Addition of anti-mouse IL-7 antibody emphasized the colony-forming ability of burst-forming unit erythroid cells. Conclusions: IL-7 produced by MS-5 cells is required for human pro-B-cell development from CD34+bone marrow cells in our culture system, and IL-7 appears to play a certain role in early human B lymphopoiesis.
UR - http://www.scopus.com/inward/record.url?scp=34548140548&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34548140548&partnerID=8YFLogxK
U2 - 10.1016/j.exphem.2007.05.019
DO - 10.1016/j.exphem.2007.05.019
M3 - Article
C2 - 17656007
AN - SCOPUS:34548140548
VL - 35
SP - 1398
EP - 1407
JO - Experimental Hematology
JF - Experimental Hematology
SN - 0301-472X
IS - 9
ER -