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Differentiation of Donor-Derived Cells Into Microglia After Umbilical Cord Blood Stem Cell Transplantation.

Takahashi K, Kakuda Y, Munemoto S, Yamazaki H, Nozaki I, Yamada M - J. Neuropathol. Exp. Neurol. (2015)

Bottom Line: Peripheral cell invasion of the brain parenchyma can only occur with disruption of the blood-brain barrier.Although the blood-brain barrier and glia limitans seemed to prevent invasion of these donor-derived cells, most of the invading donor-derived ramified cells were maintained in the cerebral cortex.This result suggests that invasion of donor-derived cells occurs through the pial membrane.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Neurology, National Hospital Organization Iou Hospital (KT); Departments of Neurology and Neurobiology of Aging (KT, YK, IN, MY) and Cellular Transplantation Biology (SM, HY), Kanazawa University Graduate School of Medical Science; and Department of Internal Medicine, Keijyu Kanazawa Hospital (SM), Kanazawa, Japan.

ABSTRACT
Recent studies have indicated that microglia originate from immature progenitors in the yolk sac. After birth, microglial populations are maintained under normal conditions via self-renewal without the need to recruit monocyte-derived microglial precursors. Peripheral cell invasion of the brain parenchyma can only occur with disruption of the blood-brain barrier. Here, we report an autopsy case of an umbilical cord blood transplant recipient in whom cells derived from the donor blood differentiated into ramified microglia in the recipient brain parenchyma. Although the blood-brain barrier and glia limitans seemed to prevent invasion of these donor-derived cells, most of the invading donor-derived ramified cells were maintained in the cerebral cortex. This result suggests that invasion of donor-derived cells occurs through the pial membrane.

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Double staining of donor-derived cells by immunohistochemistry (A, B) and in situ hybridization (C–F). (A) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti-Iba1 antibody (blue). Inset: The image at higher magnification. (B) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti–glial fibrillary acidic protein antibody (blue). Inset: The image at higher magnification. (C) In situ hybridization of the patient sample using anti-Yq12 probes (brown dot) with counterstaining of nuclei by Mayer hematoxylin solution. Arrows show Yq12-positive cells indicating donor origin. Inset: the image at higher magnification. (D) In situ hybridization of the sample from the positive control. Inset: the image at higher magnification. Arrows show Yq12-positive cells. (E) In situ hybridization of the sample from the negative control. (F) Double staining of donor-derived cells using anti-Yq12 probes (brown dot) and anti-Iba antibody (blue). Arrows show double-positive cells indicating donor origin; arrowheads show Iba1-positive and Yq12-negative cells indicating host origin. Inset: the image at higher magnification. Scale bar = 100 μm.
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Figure 2: Double staining of donor-derived cells by immunohistochemistry (A, B) and in situ hybridization (C–F). (A) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti-Iba1 antibody (blue). Inset: The image at higher magnification. (B) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti–glial fibrillary acidic protein antibody (blue). Inset: The image at higher magnification. (C) In situ hybridization of the patient sample using anti-Yq12 probes (brown dot) with counterstaining of nuclei by Mayer hematoxylin solution. Arrows show Yq12-positive cells indicating donor origin. Inset: the image at higher magnification. (D) In situ hybridization of the sample from the positive control. Inset: the image at higher magnification. Arrows show Yq12-positive cells. (E) In situ hybridization of the sample from the negative control. (F) Double staining of donor-derived cells using anti-Yq12 probes (brown dot) and anti-Iba antibody (blue). Arrows show double-positive cells indicating donor origin; arrowheads show Iba1-positive and Yq12-negative cells indicating host origin. Inset: the image at higher magnification. Scale bar = 100 μm.

Mentions: We next performed double staining to investigate the differentiation of graft cells into microglia or astrocytes. HLA-A2–positive cells stained positive for Iba1, which is representative of microglia (Fig. 2A), but not for glial fibrillary acidic protein, which is specific for astrocytes (Fig. 2B). Because these results suggested that graft cells differentiated into microglia, we also performed in situ hybridization with anti-Yq12 probes. Although most of the cells in the male positive control sample showed a brown dot in their nuclei, whereas cells in the female negative control sample had no brown dot in their nuclei, only a few cells in the transplant recipient sample had a brown dot in their nuclei, indicating Y chromosome–positive cells from the male donor (Figs. 2C–E). Moreover, in situ hybridization with anti-Yq12 probes after immunohistochemistry with anti-Iba1 antibodies showed that Y chromosome–positive cells from the male donor were Iba1-positive (Fig. 2F).


Differentiation of Donor-Derived Cells Into Microglia After Umbilical Cord Blood Stem Cell Transplantation.

Takahashi K, Kakuda Y, Munemoto S, Yamazaki H, Nozaki I, Yamada M - J. Neuropathol. Exp. Neurol. (2015)

Double staining of donor-derived cells by immunohistochemistry (A, B) and in situ hybridization (C–F). (A) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti-Iba1 antibody (blue). Inset: The image at higher magnification. (B) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti–glial fibrillary acidic protein antibody (blue). Inset: The image at higher magnification. (C) In situ hybridization of the patient sample using anti-Yq12 probes (brown dot) with counterstaining of nuclei by Mayer hematoxylin solution. Arrows show Yq12-positive cells indicating donor origin. Inset: the image at higher magnification. (D) In situ hybridization of the sample from the positive control. Inset: the image at higher magnification. Arrows show Yq12-positive cells. (E) In situ hybridization of the sample from the negative control. (F) Double staining of donor-derived cells using anti-Yq12 probes (brown dot) and anti-Iba antibody (blue). Arrows show double-positive cells indicating donor origin; arrowheads show Iba1-positive and Yq12-negative cells indicating host origin. Inset: the image at higher magnification. Scale bar = 100 μm.
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Related In: Results  -  Collection

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Figure 2: Double staining of donor-derived cells by immunohistochemistry (A, B) and in situ hybridization (C–F). (A) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti-Iba1 antibody (blue). Inset: The image at higher magnification. (B) Double staining of donor-derived cells with anti–HLA-A2 antibody (brown) and anti–glial fibrillary acidic protein antibody (blue). Inset: The image at higher magnification. (C) In situ hybridization of the patient sample using anti-Yq12 probes (brown dot) with counterstaining of nuclei by Mayer hematoxylin solution. Arrows show Yq12-positive cells indicating donor origin. Inset: the image at higher magnification. (D) In situ hybridization of the sample from the positive control. Inset: the image at higher magnification. Arrows show Yq12-positive cells. (E) In situ hybridization of the sample from the negative control. (F) Double staining of donor-derived cells using anti-Yq12 probes (brown dot) and anti-Iba antibody (blue). Arrows show double-positive cells indicating donor origin; arrowheads show Iba1-positive and Yq12-negative cells indicating host origin. Inset: the image at higher magnification. Scale bar = 100 μm.
Mentions: We next performed double staining to investigate the differentiation of graft cells into microglia or astrocytes. HLA-A2–positive cells stained positive for Iba1, which is representative of microglia (Fig. 2A), but not for glial fibrillary acidic protein, which is specific for astrocytes (Fig. 2B). Because these results suggested that graft cells differentiated into microglia, we also performed in situ hybridization with anti-Yq12 probes. Although most of the cells in the male positive control sample showed a brown dot in their nuclei, whereas cells in the female negative control sample had no brown dot in their nuclei, only a few cells in the transplant recipient sample had a brown dot in their nuclei, indicating Y chromosome–positive cells from the male donor (Figs. 2C–E). Moreover, in situ hybridization with anti-Yq12 probes after immunohistochemistry with anti-Iba1 antibodies showed that Y chromosome–positive cells from the male donor were Iba1-positive (Fig. 2F).

Bottom Line: Peripheral cell invasion of the brain parenchyma can only occur with disruption of the blood-brain barrier.Although the blood-brain barrier and glia limitans seemed to prevent invasion of these donor-derived cells, most of the invading donor-derived ramified cells were maintained in the cerebral cortex.This result suggests that invasion of donor-derived cells occurs through the pial membrane.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Neurology, National Hospital Organization Iou Hospital (KT); Departments of Neurology and Neurobiology of Aging (KT, YK, IN, MY) and Cellular Transplantation Biology (SM, HY), Kanazawa University Graduate School of Medical Science; and Department of Internal Medicine, Keijyu Kanazawa Hospital (SM), Kanazawa, Japan.

ABSTRACT
Recent studies have indicated that microglia originate from immature progenitors in the yolk sac. After birth, microglial populations are maintained under normal conditions via self-renewal without the need to recruit monocyte-derived microglial precursors. Peripheral cell invasion of the brain parenchyma can only occur with disruption of the blood-brain barrier. Here, we report an autopsy case of an umbilical cord blood transplant recipient in whom cells derived from the donor blood differentiated into ramified microglia in the recipient brain parenchyma. Although the blood-brain barrier and glia limitans seemed to prevent invasion of these donor-derived cells, most of the invading donor-derived ramified cells were maintained in the cerebral cortex. This result suggests that invasion of donor-derived cells occurs through the pial membrane.

Show MeSH
Related in: MedlinePlus