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doi: 10.1242/10.1242/jcs.00060

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Hair follicle dermal cells repopulate the mouse haematopoietic system

Department of Biological Sciences, South Road, University of Durham, Durham DH1 3LE, UK

View larger version (35K): [in a new window]   Fig. 1. Schematic presentation of the hair follicle. Black lines indicate the plane of dissection used to separate the end bulb, medium and top section of the vibrissa (see Fig. 2).

View larger version (78K): [in a new window]   Fig. 2. Follicle tissues display haematopoietic activity in vitro. CFU-GEMM assay results show colony formation from the end bulb of the mouse vibrissa (A), the medium section of the mouse vibrissa (B), the top section of the mouse vibrissa (C), the dermal sheath dissected from the mouse end bulb (D), the dermal papilla dissected from the mouse end bulb (E) and the epithelium dissected from the mouse end bulb (F). Haemoglobinised cells produced in the assay are indicated by arrows.

View larger version (74K): [in a new window]   Fig. 3. Mouse and rat follicle tissues display haematopoietic activity in vitro. CFU-A assay results from proximal tip of rat vibrissa follicle (A); dermal sheath of rat vibrissa follicle (B); dermal papilla of rat vibrissa follicle (C); proximal tip of mouse vibrissa follicle (D); dermal sheath of mouse vibrissa follicle (E); and dermal papilla of mouse vibrissa follicle (F).

View larger version (67K): [in a new window]   Fig. 4. DP and DS cultured cells display haematopoietic activity in vitro. DS and DP explants were cultured in vitro (A and B, respectively) and subjected to CFU-A assay (C and D, respectively). For comparison, E shows a CFU-A colony formed following plating of bone marrow cells.

View larger version (25K): [in a new window]   Fig. 5. (A) PCR analysis shows that cells from the follicle dermis reconstitute bone marrow, spleen and peripheral blood of recipients. The lacZ gene was detected in the peripheral blood, bone marrow and spleen of the Zin40 donor mouse, but not in the control recipients (Balb/c). M15 and M16 were transplanted with 2x104 Zin40 dermal sheath cultured cells and 2x105 Balb/c bone marrow cells. M18 and M28 were transplanted with 2x104 Zin40 dermal papilla cultured cells and 2x105 Balb/c bone marrow cells. NC, no DNA control. (B) Dermal-derived cells contribute to all haematopoietic lineages studied: confirmation by PCR. (A) Splenocytes from Balb/c recipient animals transplanted with dermal cells derived from LacZ transgenic animals (Zin40) were sorted into subpopulations (total leukocytes, CD45+; non-haematopoietic nucleated cells, CD45-; B cells, B220+; T cells, CD4+/CD8a+; and myeloid cells, CD11b and Ly6) and presence of transgenic leukocytes determined by PCR. A second PCR was performed with Wnt8b primers as a loading control. (C) Detection of transgenic donor cells by PCR. Mixtures of splenocytes from transgenic and non-transgenic animals were mixed in differing ratios from 100% transgenic to 0% transgenic and subjected to PCR for LacZ and Wnt8b.