Lab Anim Res.  2014 Mar;30(1):14-20.

Bone marrow stem/progenitor cell mobilization in C57BL/6J and BALB/c mice

• Lee H
• Che JH
• Oh JE
• Chung SS
• Jung HS
• Park KS

Abstract

Bone marrow (BM) has been considered as a reservoir of stem/progenitor cells which are able to differentiate into ectodermal, endodermal, and mesodermal origins in vitro as well as in vivo. Following adequate stimulation, such as granulocyte stimulating factor (G-CSF) or AMD3100, BM resident stem/progenitor cells (BMSPCs) can be mobilized to peripheral blood. Several host-related factors are known to participate in this mobilization process. In fact, a significant number of donors are resistant to G-CSF induced mobilization protocols. AMD3100 is currently used in combination with G-CSF. However, information regarding host-related factors which may influence the AMD3100 directed mobilization is extremely limited. In this study, we were to get some more knowledge on the host-related factors that affect the efficiency of AMD3100 induced mobilization by employing in vivo mobilization experiments. As a result, we found that C57BL/6J mice are more sensitive to AMD3100 but less sensitive to G-CSF which promotes the proliferation of BMSPCs. We excluded S1P as one of the host related factor which influences AMD3100 directed mobilization because pre-treatment of S1P receptor antagonist FTY720 did not inhibit BMSPC mobilization. Further in vitro experiments revealed that BALB/c mice, compared to C57BL/6J mice, have less BMSPCs which migrate in response to host related factors such as sphingosine-1-phosphate (S1P) and to CXCL12. We conclude that AMD3100-directed mobilization depends on the number of BMSPCs rather than on the host-related factors. These results suggest that the combination of AMD3100 and G-CSF is co-operative and is optimal for the mobilization of BMSPCs.

Keyword

Bone marrow stem/progenitor cells; Mobilization; AMD3100; G-CSF; Host-related factors

MeSH Terms

Animals
Bone Marrow*
Ectoderm
Endoderm
Granulocyte Colony-Stimulating Factor
Granulocytes
Humans
Mesoderm
Mice*
Receptors, Lysosphingolipid
Tissue Donors
Fingolimod Hydrochloride
Granulocyte Colony-Stimulating Factor
Receptors, Lysosphingolipid

Figure

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  Figure 1 BALB/c mice are poor mobilizer to AMD mobilization protocol. Age- and sex-matched C57BL/6J and BALB/c mice were mobilized with AMD3100 (5 mg/kg s.c.) for 1 h. The numbers of circulating white blood cells (A-C) and CFU-M clonogenic cells (D) per mililiter of PB are depicted. Data (mean±SD) show representative results from three independent experiments with five animals per group. *P<0.05 between C57BL/6J and BALB/c mice. CTL, counts in steady state condition; AMD3100, counts after AMD3100 induced mobilization; WBC, white blood cells; Gra, granulocytes; Lymp, lymphocytes; CFU-M, colony forming unit macrophages.

• 

  Figure 2 BALB/c mice are easy mobilizer to G-CSF mobilization protocol. Age- and sex-matched C57BL/6J and BALB/c mice were mobilized with G-CSF for 6 days. The numbers of circulating white blood cells (A-C) and CFU-M clonogenic cells (D) per mililiter of peripheral blood (PB) are shown. Data (mean±SD) show representative results from three independent experiments with five animals per group. *P<0.05 between C57BL/6J and BALB/c mice. CTL, counts in steady state condition; AMD3100, counts after AMD3100 induced mobilization; WBC, white blood cells; Gra, granulocytes; Lymp, lymphocytes; CFU-M, colony forming unit macrophages.

• 

  Figure 3 Plasma S1P does not affect the mobilization of BMSPCs in the AMD3100 protocol. Normal (PBS treated) or FTY720 (1.5 mg/kg i.p.) pre-treated C57BL/6J mice were mobilized with AMD3100 for 1 h and the numbers of circulating white blood cells (A-C) and CFU-M clonogenic cells (D) per mililiter of peripheral blood (PB) are shown. Data (mean±SD) show representative results from three independent experiments with five animals per group. *P<0.05 between normal and FTY720 pre-treated C57BL/6J mice. Closed bars, PBS pretreated; Bars with strip, FTY720 pre-treated; PBS, counts in PBS pre-treated mice after AMD3100 mobilization; FTY720, counts in FTY720 pre-treated mice after AMD3100 mobilization; WBC, white blood cells; Gra, granulocytes; Lymp, lymphocytes; CFU-M, colony forming unit macrophages.

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  Figure 4 BALB/c mice have fewer BMSPCs than C57BL/6J. The migration of BALB/c derived BMSPCs in response to S1P (100 nM) gradient (A) or in response to CXCL12 (50 ng/mL) gradient (B) are less than that of C57BL/6J. Values are the percent (%) of migrated BMSPCs compared to each input of BMSPCs (100%). The data (mean±SD) shown represent the combined results from three independent experiments carried out in triplicate per group. *P<0.05 between C57BL/6J and BALB/c mice.

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