Transplantation of HSCs and hematopoietic progenitors.

<p>(A) LSKs or CD34⁻LSKs were purified by FACS from Lin^−/lowc-Kit⁺ BM fraction of GFP mice. GFP and Lineage+propidium iodide (PI) expression of unfractionated BM cells is shown as a control. (B) Total myeloid progenitors (My-P) and CLPs were purified by FACS from Lin^−/lowThy1.2^−/lowMNCs of GFP mice. (C-E) Correlational analyses between injected cell numbers and the numbers of GFP⁺ cardiomyocytes per a recipient mouse in recipients transplanted with Lin^−/lowSca-1⁺ cells (C), LSKs (D), and CD34⁻LSKs (E). (F) Comparison of the number of GFP⁺ cardiomyocytes at the same injected cell dose in Lin^−/lowSca-1⁺ cells, LSKs, and CD34⁻LSKs recipients. The means of the number of GFP⁺ cardiomyocytes in recipients transplanted with each injected cell number are plotted. In the transplantation of LSKs, injected cell number of 1–1.8×10⁵ cells is plotted at 10⁵, that of 1–4×10⁴ cells is plotted at 10⁴, and that of 1–2×10³ cells is plotted at 10³. The greater cardiomyogenic ability existed in CD34⁻LSKs than LSKs, and in LSKs than Lin^−/lowSca-1⁺ cells.</p

Characterization of donor BM-derived GFP⁺ cells in injured heart.

<p>(A) Representative image of CD11b-expressing GFP⁺ myeloid cells. GFP⁺ hematopoietic cells in recipient cardiac tissue appeared in small and round shape. Cardiac section was stained with anti-CD11b (red, Cy3) and DAPI (blue). Inset: high magnification of GFP⁺CD11b⁺ cells. (B) Representative image of a vimentin-expressing GFP⁺ fibroblast. Cardiac section was stained with anti-vimentin (red, Cy3). The fibroblast was present adjacent to striated cardiomyocytes in differential interference contrast (DIC) image. (C and D) Representative images of a TnI- (C) or Cx43- (D) expressing GFP⁺ striated cardiomyocyte. Cardiac sections were stained with anti-TnI (C; red, Cy3), anti-Cx43 (D; yellow, Cy5), and DAPI (blue). Cardiac sections are from recipients transplanted with unfractionated BM cells. All merged images were obtained from the same confocal plane. Scale bars = 50 µm, (A)-inset 10 µm.</p

The number of donor-derived cardiomyocytes after transplantation of purified BM cells.

a<p>P = 0.0095 by Mann-Whitney U test, Lin^−/lowCD45⁺ versus Lin^−/lowCD45^−.</p>b<p>P = 0.0021 by Mann-Whitney U test, Myeloid progenitor versus CLP.</p>c<p>P = 0.0004 by Mann-Whitney U test, CMP versus CLP.</p>d<p>P = 0.1142 by Mann-Whitney U test, Myeloid progenitor versus CLP.</p>*<p>GFP⁺ cardiomyocytes were counted in 40 contiguous sections from apex of the heart per a mouse. Detailed information of histological analysis is described in Materials and Methods, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0062506#pone.0062506.s008" target="_blank">Materials and Methods S1</a>.</p><p>Abbreviations. BM: bone marrow, PB: peripheral blood, HSC: hematopoietic stem cell, LSK: Lin⁻Sca-1⁺c-Kit⁺, CMP: common myeloid progenitor, GMP: granulocyte/monocyte progenitor, CLP: common lymphoid progenitor.</p