Detection of bone marrow–derived lung epithelial cells (original) (raw)
Related papers
Failure of Bone Marrow to Reconstitute Lung Epithelium
American Journal of Respiratory Cell and Molecular Biology, 2005
A new paradigm of epithelial tissue reconstitution has been suggested whereby circulating cells derived from bone marrow contribute to a variety of epithelial cell types. With regard to the lung, several recent reports have used immunofluorescence microscopy to demonstrate engraftment of bone marrow-derived cells as type II pneumocytes, the endogenous progenitors of the lung alveolus. We show here that immunofluorescence microscopy, as has been used in previous reports, cannot reliably identify rare engrafted cells in lung tissue sections after transplantation of bone marrow cells or purified hematopoietic stem cells tracked with ubiquitous labels. We have employed a lineage-specific reporter system based on transgenic mice that express the GFP reporter gene only in lung epithelial cells (surfactant protein C-GFP) to assay for engrafted cells by flow cytometry, histology, and molecular methods. Using this approach to evaluate transplant recipients, including those subjected to bleomycin-induced lung injury, we demonstrate that when autofluorescence, dead cells, and contaminating blood cells are excluded from analysis, there is no detectable reconstitution of lung alveolar epithelial cells by unfractionated bone marrow cells or purified hematopoietic stem cells.
Bone marrow-derived cells as progenitors of lung alveolar epithelium
Development, 2001
We assessed the capacity of plastic-adherent cultured bone marrow cells to serve as precursors of differentiated parenchymal cells of the lung. By intravenously delivering lacZ-labeled cells into wild-type recipient mice after bleomycin-induced lung injury, we detected marrow-derived cells engrafted in recipient lung parenchyma as cells with the morphological and molecular phenotype of type I pneumocytes of the alveolar epithelium. At no time after marrow cell injection, did we detect any engraftment as type II pneumocytes. In addition, we found that cultured and fresh aspirates of bone marrow cells can express the type I pneumocyte markers, T1α and aquaporin-5. These observations challenge the current belief that adult alveolar type I epithelial cells invariably arise from local precursor cells and raise the possibility of using injected marrow-derived cells for therapy of lung diseases characterized by extensive alveolar damage.
Evidence that Bone Marrow Cells Do Not Contribute to the Alveolar Epithelium
2005
An ongoing controversy is the role of marrow cells in populating the alveolar epithelium. In this study, we employed flow cytometry and histologic techniques to evaluate this process. Donor bone marrow was harvested from transgenic mice expressing the LacZ or eGFP gene ubiquitously, or under the control of the human surfactant protein (SP)-C promoter, and transplanted into lethally irradiated, neonatal mice. In recipients transplanted with marrow that express eGFP or lacZ ubiquitously, light microscopy revealed cells whose morphology and location were compatible with a type II cell phenotype. Consistent with this, fluorescent microscopy suggested colocalization of eGFP and pro-SP-C proteins in single cells. In mice transplanted with SP-C-eGFP marrow, engraftment was not detectable by histology or flow cytometry. We therefore used deconvolution microscopy to reanalyze histologic sections that were thought to show marrow-derived type II cells. We found that all putative marrow-derived pneumocytes resulted from the overlapping fluorescent signals of an endogenous pro-SP-Cϩ type II cell and a donor-derived eGFPϩ cell. Taken together, our observations underscore the technical difficulties associated with evaluating engraftment in lung, and argue against a contributory role for marrow cells in populating the alveolar epithelium.
Nonhematopoietic Cells are the Primary Source of Bone Marrow-Derived Lung Epithelial Cells
STEM CELLS, 2012
Previous studies have demonstrated that bone marrow (BM)-derived cells differentiate into nonhematopoietic cells of multiple tissues. To date, it remains unknown which population(s) of BM cells are primarily responsible for this engraftment. To test the hypothesis that nonhematopoietic stem cells in the BM are the primary source of marrow-derived lung epithelial cells, either wild-type hematopoietic or nonhematopoietic BM cells were transplanted into irradiated surfactant-protein-C (SPC)-null mice. Donor-derived, SPC-positive type 2 pneumocytes were predominantly detected in the lungs of mice receiving purified nonhematopoietic cells and were absent from mice receiving purified hematopoietic stem and progenitor cells. We conclude that cells contained in the nonhematopoietic fraction of the BM are the primary source of marrow-derived lung epithelial cells. These nonhematopoietic cells may represent a primitive stem cell population residing in adult BM. STEM CELLS 2012;30:491-499
The Journal of Heart and Lung Transplantation, 2009
Background: There is a growing expectation that cell-based therapies will prove effective for a wide range of conditions including lung diseases such as cystic fibrosis. The promise of these therapies will depend largely on effective delivery and engraftment. In this study, in the setting of human lung transplantation, we sought to determine whether exogenous epithelial cells are able to engraft the transplanted organ and if cells of a similar phenotype could be detected in peripheral blood.
Differentiation of bone marrow cells in culture and in vivo
International Congress Series, 2003
Recently, the bone marrow has been demonstrated to include progenitor cells that give rise to a large variety of cell types. We found that 3.5 -4% of adult rat bone marrow cells express hepatocyte growth factor (HGF) receptor (c-Met) and a-fetoprotein (AFP). Using a hepatocyte growth medium supplemented with hepatocyte growth factor and epidermal growth factor, we succeeded in inducing hepatocyte-like cells from the bone marrow cells in culture. The cells expressed albumin as shown by immunocytochemistry and mRNAs of tryptophan-2,3-dioxygenase and tyrosine aminotransferase, the late markers of mature hepatocytes. Furthermore, in a skin reconstitution system in vivo, we showed that mouse bone marrow cells differentiated to several different cell types of skin within 3 weeks when combined with mouse embryonic skin cells.
Bone Marrow-Derived Cells Contribute to Epithelial Engraftment during Wound Healing
American Journal of Pathology, 2004
Recent findings suggest that bone marrow-derived cells (BMDC) may contribute to tissue maintenance throughout the body. However, it is not yet known whether marrow-derived epithelial cells are capable of undergoing proliferation. Our laboratory has shown that BMDC engraft as keratinocytes in the skin at low levels (< 1%) in the absence of injury. Here we show that skin damage affects the degree of engraftment of BMDC as keratinocytes and that the keratinocytes are actively cycling. Female mice reconstituted with sex-mismatched BM were wounded by punch biopsy and incision. At the wound site, engraftment of BMDC as epidermal cells increased within 1 day, and continued to increase to approximately 4% by 3 weeks after injury. Using a Cre-lox system, fusion of BMDC with epithelial cells was ruled out. BMDC-derived epithelial cells at the wound edges expressed Ki67, a marker for actively cycling cells, and this proliferation correlated with an increase in the number of donor-derived cells within the wound. Donorderived cytokeratin 5-expressing cells were rare, suggesting that BMDC do not engraft as epidermal stem cells, and the level of engraftment peaked and then decreased over time, further suggesting that BMDC may assist in early wound healing by engrafting as transit-amplifying cells, which then differentiate into keratinocytes.
Tissue Engineering, 2007
Mesenchymal stem cells (MSCs) from bone marrow are a potential source for reconstructive therapy. In vitro, MSCs differentiate into cells of mesodermal and ectodermal lineages but rarely into cells of endodermal lineage. We developed an in vitro model to study the endodermal differentiation of MSCs using co-culture of MSCs and transformed lung epithelial (A-549) cells. The cells were separated using a cellimpermeable membrane to eliminate the possibility of cell fusion. Under these conditions, MSCs expressed several lung epithelial markers (cytokeratins 5, 8, 14, 18, 19, pro-surfactant protein C, zonula occludens-1), detected using quantitative reverse transcriptase polymerase chain reaction and Western blot, and b-catenin signaling was activated in MSCs. Treatment of MSCs with 10 to 20 mM lithium chloride activated the b-catenin pathway and enhanced expression of epithelial markers, although this activation was transient. We conclude that A-549 cells can trigger epithelial differentiation of MSCs by a paracrine mechanism that may include activation of b-catenin signaling.
Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell
Cell, 2001
is not yet known. Here we study whether a unique bone marrow subpopulation highly enriched for hematopoi-HSC are present in mouse bone marrow at a frequency Yale University School of Medicine of 1 in 10 5 cells ). The rarity of these New Haven, Connecticut 06520 cells and the absence of specific markers have made 3 Department of Pathology the search for a pure HSC population a challenge for New York University Medical School the past 50 years. The lack of ideal in vitro assays for New York, New York 10016 HSC requires that functional assays be utilized to estab-4 Oncology Center lish their presence. We and others have shown that LTR Johns Hopkins School of Medicine is possible with small numbers (1-10) of HSC (Jones et Baltimore, Maryland 21231 al., 1996; Spangrude et al., 1995; Osawa et al., 1996), but serial transplantation (self-renewal) of single cell reconstituted recipients serving as donors for new recipi-Summary ents has not yet been shown convincingly. We test, using a two day homing protocol, whether Purification of rare hematopoietic stem cell(s) (HSC)