- Email: [email protected]
Derivation of lung epithelium from bone marrow cells
ISeT
Society for Cellular TheraP~
--Cytotherapy (2003) Vol. 5, No. 2, 169-173
(!aylor&Francis • healthsciences
Derivation of lung epithelium from bone marrow
cells DN Kotton and A Fine The Pulmonary Center, Boston University School ofMedicine, MA, USA
Recent studies demonstrate the capacity ofBM-derived celts to engraft
summarize our work demonstrating that cultured adherent marrow
as differentiated celts of a van'ety of organs, including lung. In this
celts can serve as progenitors oflung alveolar epithelium.
paper, we review the current state-of-the-art in this area. We also
Introduction Fresh BM culmred on plastic can be separated into two general populations: plastic-adherent and non-adherent cells. Cells that remain floating in the supernatant contain
been identified in humans [6,7) and rats [8), but have not yet been described in mice. Fibroblast-like MSCs are able to synthesize matrix proteins, such as Collagen I, Collagen IV, laminin and fibronectin [2,4,5,9).
hematopoietic stem cells (HSCs). These cells serve as precursors to circulating cells of the blood [I). In contrast, marrow cells that adhere to plastic grow into colonies of heterogeneous cells. Enriched within plastic-adherent marrow are cells referred to either as marrow stromal cells (MSCs), because they appear to arise from the complex array of supporting strucmres found in marrow, or as mesenchymal stem cells (also abbreviated MSCs), because of their ability to proliferate and differentiate into cells such as bone, cartilage, and muscle [2 - 5). Whether MSCs and HSCs originate from a single precursor is not clear at this time. Plastic-adherent BM cells consist of three discernible morphologic subtypes: • Small rounded cells posltlve for Factor VIII Ag that may be of endothelial stem-cell lineage • Small stellate cells positive for mac-I ICD II band CD45 that are likely of myeloid lineage
• Large polygonal fibroblast-like cells negative for CD34, and negative for myeloid markers [4,5). A progenitor of fibroblast-like MSCs with rapid proliferative potential, recycling stem cells (RS cells) have
Non-hematopoietic tissue derived from BM cells The in vivo ability of culmred MSCs, fresh unfractionated whole marrow, or highly enriched HSC types to form nonhematopoietic tissue has been demonstrated in a variety of publications. In most of these smdies, labeled donor BM cells are infused into marrow-ablated recipient mice. In several such experiments, precursor cells, thought to be of HSC origin, successfully served as progenitors for fully differentiated cells of liver, muscle, brain, heart, and lung, while also successfully reconstimting the recipients' ablated BM [10-16). In other experiments, cells of MSC origin differentiated into mesenchymal tissue, such as bone, cartilage, and muscle, as well as non-mesenchymal tissue, such as endothelium, hepatocytes, and neural cells after direct injection into brain [2,17- 24). Recipient marrow ablation, which facilitates and permits migration of injected cells into and through the marrow, could be a factor affecting the outcome of engraftment smdies involving BMT The importance of marrow ablation during engraftment of MSCs is not evident, however, particularly in view of
Corresp011dence to: DN Kotton, The Pulmonary Center, R-304, Boston University School of Medicine, 715 AI&any Street, Boston, MA 021182397, USA
©
2003 ISCT
001: 10.1080/14653240310001073
170
DN Kottorl arid A Firle
studies demonstrating the marked radio- and chemo-
its ability to rapidly home to an intermediate recipient's
resistance of this cell type [3].
marrow [13] .
Ability of BM cells to engraft in lung tissue After BMT, the appearance of donor marrow-derived
Engraftment of plastic-adherent marrow cells as Type I pneumocytes
collagen-producing cells in lung tissue occurs in a time-
We found that cultured plastic-adherent marrow cells from
dependent process [19,20] . The role of pre-BMT TBI, a
lacZ + donor mice can engraft in bleomycin-injured
known inducer of lung injury, in mediating these observa-
recipient lungs as Type I cells of the alveolar epithelium
tions is uncertain. Histologic analysis of recipient murine
[26]. Through X-gal staining of lacZ + cells, we identified
lungs 1- 20 weeks after injection of labeled donor marrow
donor-derived cells in recipient lungs with the typical
.. , cells found donor-derived cells in the lung parenchyma '..:"[ 19,25]. Most notably, Krause et al. showed widespread
Type I alveolar cell morphology (Figure 1). These engrafted cells express the Type I cell apical membrane
multi-lineage engraftment as epithelial cells in various
protein, Tlrx [26]. Analysis at early time points (1- 5 days)
organs, including lung alveolar Type 11 and bronchial cells
after injection [26] demonstrated no evidence of engraft-
11 months after transplantation of a single BM stem cell.
ment as Type 11 pneumocytes, the endogenous progenitor
This cell was functionally purified from donor marrow by
of Type I cells [27] .
Figure 1. Paraffin and plastic sections of X-gal-stained mouse lungs 30 days after cell injection. (A) Low magnification (40 x) and (B) high magnification (1000 x ) views of a representative recipient lung showing engrafted donor-derived cells as thin, flat blue cells consistent with a Type I pneumocyte morphology. (e) For comparison, an X-gal-stained negative control lung (lacZ-negative marrow injected into a lacZ-negative recipient) shows no blue cells. (D) A thin plastic section (2 ).lm) of recipient mouse lung confirms the Type I pneumocyte morphology of engrafted donor-dm'ved cells. Sections are counter-stained with Nuclear Fast Red. Bars, 200 m (A); 30 m (B- Dj.
Marrow-derived lung epithelium
171
Figure 2. Histological analyses of adherem BM cultures. (A) Three cell morphologies are discerrtible: larger polygonal fibroblast-like cells (black arrow), smaller stellate myeloid cells (red arrow), and small round ertdothelial-like cells (purple arrow). (H- D) Immtmostaining of cultures with ami-mouse Tla Ab (brown staining; H, arrows). (C) At higher magnification, the T l a -positive cell demonstrates fibroblast-like morphology. (D) Staining within a cell colony is heterogerteous. Bars, 20 m (A); 200 m (H); 15 m (C); 25 m (D). (A)
C ultu red
T l a SPC AQ5
T la SPC AQ5
CC-lO (B)
Figure 3. (A). Nested RT-PCR analysis for lung epithelial markers in plastic-adherertt cultured marrow cells (7 days) and fresh BM. Positive expression of T la and aquaporirt-5 (AQJ) was rtoted urtder both conditions. Surfactant protein-C (SPC) was not expressed. (H). RT-PCR analysis showing expression of the airway Clara cell marker (CC 10) ifI plastic-adherent marrow cultured for 6 afld 13 days. Lung served as a positive contro/.
172
DN Kaftan and A Fine
replacement of specific cell types damaged or lost during disease. The identification of specific marrow cell type(s) that can assume lung cell phenotypes, and the characterization of mechanisms controlling this differentiation process are two of the many challenges facing this nascent field.
References Spangrude Gj, Heimfeld S, Weissman IL. Purification and characterization of mouse hematopoietic stem cells [published erratum
Figure 4. RT-PCR analysis for expression of transcription factors, TTF-l and HNF-3P in fresh (Fr) or cultured (Cx'd) BM cells. PCR
appears
In
Science
1989;244:1030].
2
oflung cDNA was used as a positive control.
Prockop DJ. Marrow stromal cells as stem cells for non hem atopoietic tissues. Seimee 1997;276:71-4. Bianco P, Robey PG. Marrow stromal stem cells.
Lung epithelial gene expression in plasticadherent cell cultures We sought to identify expression oflung epithelial genes in cultured plastic-adherent marrow cells. TIel, an apical membrane protein of unknown function, is highly specific in the lung for Type I alveolar cells [28- 32]. Approximately 10% of plastic-adherent marrow cells are TIel positive after 1 week in culture (Figure 2B). TIel positive cells have the morphological features of the polygonal fibrobast-like MSC cell sub-population. Expression of TIel within a clone of the polygonal fibroblast-like cells is heterogeneous (Figure 20) [26]. Aquaporin-5, another Type I cell marker, was found by RT-PCR to be expressed in marrow preparations (Figure 3A). In contrast, we found that the alveolar Type 11 cellspecific marker, surfactant protein-C (SP-C) was not expressed in fresh or cultured cells (Figure 3A) [26]. In addition, we found that the message for CC-10, an airway epithelial marker (Figure 3B) was expressed in cultured, but not fresh marrow. We also utilized RT-PCR to assess expression of the transcription factors, TTF-1 and HNF3p. These molecules regulate key transcriptional events in airway and alveolar epithelial differentiation [33,34]. We found that fresh and cultured BM cells express these genes (Figure 4).
Seimee
1989;241:58-62.
J Cli11 luvest
2000; 105: 1663 - 8.
4
Phinney DG, Kopen G, Isaacson RL, Prockop DJ. Plastic adherent stromal cells from the bone marrow of commonly used strains of inbred mice: variations in yield, growth, and differentiation.
J Cell Bioehem
1999;72:570-85.
Pirrenger MF, Mackay AM, Beck SC et al. Multilineage potential of adult human mesenchymal stem cells. Seimee 1999)84:143-7.
6
Colter DC, Sekiya I, Prockop DJ. Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proe Natl Aead Sei
USA 2001;98:7841- 5. 7
Colter DC, Class R, DiGirolamo CM, Prockop DJ. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proe Nat! Aead Sci USA 2000;97:3213- 8.
8 javazon EH, Colter DC, Schwarz Ej, Prockop DJ. Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells. Stem Cells 2001;19:219-25. 9
Phinney DG, Kopen G, Righter W
et al. Donor variation in the
growth properties and osteogenic potential of human marrow stromal cells. 10
J Cell Bioebem
1999;75:424- 36.
Lagasse E, Connors H, Al Dhalimy M
et at. Purified hemato-
poietic stem cells can differentiate into hepatocytes
i11
vivo. Nat
Med 2000;6: 1229 - 34. 11
Gussoni E, Soneoka Y, Strickland CD
et al. Dystrophin
expression in the mdx mouse restored by stem cell transplantation. Nature 1999;401:390-4. 12
Eglitis MA, Mezey E. Hematopoietic cells differentiate into both microglia and macroglia in the brains of adult mice. Proe Nat!
Aead Sei USA 1997;94:4080- 5.
Conclusion The physiologic relevance of experimental data showing that marrow-derived cells can engraft as differentiated lung cells remains uncertain. Nevertheless, these observations suggest a powerful therapeutic strategy for the
13
Krause OS, Theise NO, Collector MI
et al. Multi-organ, multi-
lineage engraftment by a single bone marrow-derived stem cell.
Cell 2001;105:369-77. 14
Orlic 0, Kajstura j, Chimenti S
et al. Bone marrow cells
regenerate infarcted myocardium. Nature 2001;410:701- 5.
Marrow-derived lung epithelium
15
16
Orlic D, Kajstura J, Chimenti S et al. Mobilized bone marrow
immarure hematopoietic cells into adult mouse brain parench-
Proc Natl Acad Sci USA 2001;98:10344-9. Petersen BE, Bowen WC, Patrene KD et at. Bone marrow as a
yma under GFP-expressing bone marrow chimera. Biochem
Biophys Res Commun 1999;262:610-4. 26
cells in children with osteogenesis imperfecta [see comments].
27
entiate into astrocytes after injection into neonatal mouse brains.
Proc Natl Acad Sci USA 1999;96:10711-6. Pereira RF, Halford KW, O'Hara MD et al. Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. Proc Natl Acad Sci
USA 1995;92:4857-61. Pereira RF, O'Hara MD, Laptev AV et al. Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta.
Proc Natl Acad Sci USA 1998;95:1142-7. 21
Zhao LR, Duan WM, Reyes M et al. Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol 2002; 174: 11- 20.
22
Reyes M, Lund T, Lenvik T et al. Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 2001;98:2615-25.
23
Reyes M, Verfaillie CM. Characterization of multipotent adult progenitor cells, a subpopulation of mesenchymal stem cells. Ann
NYAcad Sci 2001;938:231-3. 24
N02. Exp Mol Pathol 1975;22:142-50.
Kopen GC, Prockop DJ, Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum, and they differ-
20
Reyes M, Dudek A, Jahagirdar B et al. Origin of endothelial progenitors in human postnatal bone marrow. 2002; I09:3 37- 46.
Evans MJ, Cabral LJ, Stephens RJ, Freeman G. Transformation of alveolar type 2 cells to type I cells following exposure to
Nat Med 1999;5:309- 13.
19
Korron DN, Ma BY, Cardoso WV et al. Bone marrow-derived cells as progenitors of lung alveolar epithelium. Development 2001;128:5181- 8.
Horwitz EM, Prockop DJ, Fitzpatrick LA et al. Transplantability and therapeutic effects of bone marrow-derived mesenchymal
18
Ono K, Takii T, Onozaki K et al. Migration of exogenous
cells repair the infarcted heart, improving function and survival.
potential source of hepatic oval cells. Science 1999;284:1168 - 70. 17
25
173
J Clin Invest
28
Williams MC, Cao Y, Hinds A et al. TI alpha protein is developmentally regulated and expressed by alveolar type I cells, choroid plexus, and ciliary epithelia of adult rats. Am
J
Respir Cell Mol BioI 1996;14:577-85. 29 Rishi AK, Joyce-Brady M, Fisher J et al. Cloning, characterization, and development expression of a rat lung alveolar type I cell gene in embryonic endodermal and neural derivatives. Dev BioI 1995;167:294- 306. 30 Danto SI, Zabski SM, Crandall ED. Reactivity of alveolar epithelial cells in primary culture with type I cell monoclonal antibodies. Am J Respir Cell Mol BioI 1992;6:296- 306. 31 Cao YX,JeanJC, Williams MC. Cytosine methylation of an Spl site contributes to organ-specific and cell-specific regulation of expression of the lung epithelial gene tlalpha. Biochem J 2000;350:883 - 90. 32 Ramirez MI, Rishi AK, Cao YX, Williams MC. TGT3, thyroid transcription factor I, and Spl elements regulate transcriptional activity of the l.3-kilobase pair promoter of TI alpha, a lung alveolar type I cell gene. J BioI Chem 1997;272:26285 - 94. 33 Costa RH, Kalinichenko VV, Lim L. Transcription factors in mouse lung development and function. Am J Physiol Lung Cell Mol Physiol 2001;280:L823-38. 34 TichelaarJW, Lim L, Costa RH, WhitserrJA. HNF-3/forkhead homologue-4 influences lung morphogenesis and respiratory epithelial cell differentiation if1 vivo. Dev BioI 1999;213:405 -17.
Society for Cellular TheraP~
--Cytotherapy (2003) Vol. 5, No. 2, 169-173
(!aylor&Francis • healthsciences
Derivation of lung epithelium from bone marrow
cells DN Kotton and A Fine The Pulmonary Center, Boston University School ofMedicine, MA, USA
Recent studies demonstrate the capacity ofBM-derived celts to engraft
summarize our work demonstrating that cultured adherent marrow
as differentiated celts of a van'ety of organs, including lung. In this
celts can serve as progenitors oflung alveolar epithelium.
paper, we review the current state-of-the-art in this area. We also
Introduction Fresh BM culmred on plastic can be separated into two general populations: plastic-adherent and non-adherent cells. Cells that remain floating in the supernatant contain
been identified in humans [6,7) and rats [8), but have not yet been described in mice. Fibroblast-like MSCs are able to synthesize matrix proteins, such as Collagen I, Collagen IV, laminin and fibronectin [2,4,5,9).
hematopoietic stem cells (HSCs). These cells serve as precursors to circulating cells of the blood [I). In contrast, marrow cells that adhere to plastic grow into colonies of heterogeneous cells. Enriched within plastic-adherent marrow are cells referred to either as marrow stromal cells (MSCs), because they appear to arise from the complex array of supporting strucmres found in marrow, or as mesenchymal stem cells (also abbreviated MSCs), because of their ability to proliferate and differentiate into cells such as bone, cartilage, and muscle [2 - 5). Whether MSCs and HSCs originate from a single precursor is not clear at this time. Plastic-adherent BM cells consist of three discernible morphologic subtypes: • Small rounded cells posltlve for Factor VIII Ag that may be of endothelial stem-cell lineage • Small stellate cells positive for mac-I ICD II band CD45 that are likely of myeloid lineage
• Large polygonal fibroblast-like cells negative for CD34, and negative for myeloid markers [4,5). A progenitor of fibroblast-like MSCs with rapid proliferative potential, recycling stem cells (RS cells) have
Non-hematopoietic tissue derived from BM cells The in vivo ability of culmred MSCs, fresh unfractionated whole marrow, or highly enriched HSC types to form nonhematopoietic tissue has been demonstrated in a variety of publications. In most of these smdies, labeled donor BM cells are infused into marrow-ablated recipient mice. In several such experiments, precursor cells, thought to be of HSC origin, successfully served as progenitors for fully differentiated cells of liver, muscle, brain, heart, and lung, while also successfully reconstimting the recipients' ablated BM [10-16). In other experiments, cells of MSC origin differentiated into mesenchymal tissue, such as bone, cartilage, and muscle, as well as non-mesenchymal tissue, such as endothelium, hepatocytes, and neural cells after direct injection into brain [2,17- 24). Recipient marrow ablation, which facilitates and permits migration of injected cells into and through the marrow, could be a factor affecting the outcome of engraftment smdies involving BMT The importance of marrow ablation during engraftment of MSCs is not evident, however, particularly in view of
Corresp011dence to: DN Kotton, The Pulmonary Center, R-304, Boston University School of Medicine, 715 AI&any Street, Boston, MA 021182397, USA
©
2003 ISCT
001: 10.1080/14653240310001073
170
DN Kottorl arid A Firle
studies demonstrating the marked radio- and chemo-
its ability to rapidly home to an intermediate recipient's
resistance of this cell type [3].
marrow [13] .
Ability of BM cells to engraft in lung tissue After BMT, the appearance of donor marrow-derived
Engraftment of plastic-adherent marrow cells as Type I pneumocytes
collagen-producing cells in lung tissue occurs in a time-
We found that cultured plastic-adherent marrow cells from
dependent process [19,20] . The role of pre-BMT TBI, a
lacZ + donor mice can engraft in bleomycin-injured
known inducer of lung injury, in mediating these observa-
recipient lungs as Type I cells of the alveolar epithelium
tions is uncertain. Histologic analysis of recipient murine
[26]. Through X-gal staining of lacZ + cells, we identified
lungs 1- 20 weeks after injection of labeled donor marrow
donor-derived cells in recipient lungs with the typical
.. , cells found donor-derived cells in the lung parenchyma '..:"[ 19,25]. Most notably, Krause et al. showed widespread
Type I alveolar cell morphology (Figure 1). These engrafted cells express the Type I cell apical membrane
multi-lineage engraftment as epithelial cells in various
protein, Tlrx [26]. Analysis at early time points (1- 5 days)
organs, including lung alveolar Type 11 and bronchial cells
after injection [26] demonstrated no evidence of engraft-
11 months after transplantation of a single BM stem cell.
ment as Type 11 pneumocytes, the endogenous progenitor
This cell was functionally purified from donor marrow by
of Type I cells [27] .
Figure 1. Paraffin and plastic sections of X-gal-stained mouse lungs 30 days after cell injection. (A) Low magnification (40 x) and (B) high magnification (1000 x ) views of a representative recipient lung showing engrafted donor-derived cells as thin, flat blue cells consistent with a Type I pneumocyte morphology. (e) For comparison, an X-gal-stained negative control lung (lacZ-negative marrow injected into a lacZ-negative recipient) shows no blue cells. (D) A thin plastic section (2 ).lm) of recipient mouse lung confirms the Type I pneumocyte morphology of engrafted donor-dm'ved cells. Sections are counter-stained with Nuclear Fast Red. Bars, 200 m (A); 30 m (B- Dj.
Marrow-derived lung epithelium
171
Figure 2. Histological analyses of adherem BM cultures. (A) Three cell morphologies are discerrtible: larger polygonal fibroblast-like cells (black arrow), smaller stellate myeloid cells (red arrow), and small round ertdothelial-like cells (purple arrow). (H- D) Immtmostaining of cultures with ami-mouse Tla Ab (brown staining; H, arrows). (C) At higher magnification, the T l a -positive cell demonstrates fibroblast-like morphology. (D) Staining within a cell colony is heterogerteous. Bars, 20 m (A); 200 m (H); 15 m (C); 25 m (D). (A)
C ultu red
T l a SPC AQ5
T la SPC AQ5
CC-lO (B)
Figure 3. (A). Nested RT-PCR analysis for lung epithelial markers in plastic-adherertt cultured marrow cells (7 days) and fresh BM. Positive expression of T la and aquaporirt-5 (AQJ) was rtoted urtder both conditions. Surfactant protein-C (SPC) was not expressed. (H). RT-PCR analysis showing expression of the airway Clara cell marker (CC 10) ifI plastic-adherent marrow cultured for 6 afld 13 days. Lung served as a positive contro/.
172
DN Kaftan and A Fine
replacement of specific cell types damaged or lost during disease. The identification of specific marrow cell type(s) that can assume lung cell phenotypes, and the characterization of mechanisms controlling this differentiation process are two of the many challenges facing this nascent field.
References Spangrude Gj, Heimfeld S, Weissman IL. Purification and characterization of mouse hematopoietic stem cells [published erratum
Figure 4. RT-PCR analysis for expression of transcription factors, TTF-l and HNF-3P in fresh (Fr) or cultured (Cx'd) BM cells. PCR
appears
In
Science
1989;244:1030].
2
oflung cDNA was used as a positive control.
Prockop DJ. Marrow stromal cells as stem cells for non hem atopoietic tissues. Seimee 1997;276:71-4. Bianco P, Robey PG. Marrow stromal stem cells.
Lung epithelial gene expression in plasticadherent cell cultures We sought to identify expression oflung epithelial genes in cultured plastic-adherent marrow cells. TIel, an apical membrane protein of unknown function, is highly specific in the lung for Type I alveolar cells [28- 32]. Approximately 10% of plastic-adherent marrow cells are TIel positive after 1 week in culture (Figure 2B). TIel positive cells have the morphological features of the polygonal fibrobast-like MSC cell sub-population. Expression of TIel within a clone of the polygonal fibroblast-like cells is heterogeneous (Figure 20) [26]. Aquaporin-5, another Type I cell marker, was found by RT-PCR to be expressed in marrow preparations (Figure 3A). In contrast, we found that the alveolar Type 11 cellspecific marker, surfactant protein-C (SP-C) was not expressed in fresh or cultured cells (Figure 3A) [26]. In addition, we found that the message for CC-10, an airway epithelial marker (Figure 3B) was expressed in cultured, but not fresh marrow. We also utilized RT-PCR to assess expression of the transcription factors, TTF-1 and HNF3p. These molecules regulate key transcriptional events in airway and alveolar epithelial differentiation [33,34]. We found that fresh and cultured BM cells express these genes (Figure 4).
Seimee
1989;241:58-62.
J Cli11 luvest
2000; 105: 1663 - 8.
4
Phinney DG, Kopen G, Isaacson RL, Prockop DJ. Plastic adherent stromal cells from the bone marrow of commonly used strains of inbred mice: variations in yield, growth, and differentiation.
J Cell Bioehem
1999;72:570-85.
Pirrenger MF, Mackay AM, Beck SC et al. Multilineage potential of adult human mesenchymal stem cells. Seimee 1999)84:143-7.
6
Colter DC, Sekiya I, Prockop DJ. Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proe Natl Aead Sei
USA 2001;98:7841- 5. 7
Colter DC, Class R, DiGirolamo CM, Prockop DJ. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proe Nat! Aead Sci USA 2000;97:3213- 8.
8 javazon EH, Colter DC, Schwarz Ej, Prockop DJ. Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells. Stem Cells 2001;19:219-25. 9
Phinney DG, Kopen G, Righter W
et al. Donor variation in the
growth properties and osteogenic potential of human marrow stromal cells. 10
J Cell Bioebem
1999;75:424- 36.
Lagasse E, Connors H, Al Dhalimy M
et at. Purified hemato-
poietic stem cells can differentiate into hepatocytes
i11
vivo. Nat
Med 2000;6: 1229 - 34. 11
Gussoni E, Soneoka Y, Strickland CD
et al. Dystrophin
expression in the mdx mouse restored by stem cell transplantation. Nature 1999;401:390-4. 12
Eglitis MA, Mezey E. Hematopoietic cells differentiate into both microglia and macroglia in the brains of adult mice. Proe Nat!
Aead Sei USA 1997;94:4080- 5.
Conclusion The physiologic relevance of experimental data showing that marrow-derived cells can engraft as differentiated lung cells remains uncertain. Nevertheless, these observations suggest a powerful therapeutic strategy for the
13
Krause OS, Theise NO, Collector MI
et al. Multi-organ, multi-
lineage engraftment by a single bone marrow-derived stem cell.
Cell 2001;105:369-77. 14
Orlic 0, Kajstura j, Chimenti S
et al. Bone marrow cells
regenerate infarcted myocardium. Nature 2001;410:701- 5.
Marrow-derived lung epithelium
15
16
Orlic D, Kajstura J, Chimenti S et al. Mobilized bone marrow
immarure hematopoietic cells into adult mouse brain parench-
Proc Natl Acad Sci USA 2001;98:10344-9. Petersen BE, Bowen WC, Patrene KD et at. Bone marrow as a
yma under GFP-expressing bone marrow chimera. Biochem
Biophys Res Commun 1999;262:610-4. 26
cells in children with osteogenesis imperfecta [see comments].
27
entiate into astrocytes after injection into neonatal mouse brains.
Proc Natl Acad Sci USA 1999;96:10711-6. Pereira RF, Halford KW, O'Hara MD et al. Cultured adherent cells from marrow can serve as long-lasting precursor cells for bone, cartilage, and lung in irradiated mice. Proc Natl Acad Sci
USA 1995;92:4857-61. Pereira RF, O'Hara MD, Laptev AV et al. Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta.
Proc Natl Acad Sci USA 1998;95:1142-7. 21
Zhao LR, Duan WM, Reyes M et al. Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol 2002; 174: 11- 20.
22
Reyes M, Lund T, Lenvik T et al. Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 2001;98:2615-25.
23
Reyes M, Verfaillie CM. Characterization of multipotent adult progenitor cells, a subpopulation of mesenchymal stem cells. Ann
NYAcad Sci 2001;938:231-3. 24
N02. Exp Mol Pathol 1975;22:142-50.
Kopen GC, Prockop DJ, Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum, and they differ-
20
Reyes M, Dudek A, Jahagirdar B et al. Origin of endothelial progenitors in human postnatal bone marrow. 2002; I09:3 37- 46.
Evans MJ, Cabral LJ, Stephens RJ, Freeman G. Transformation of alveolar type 2 cells to type I cells following exposure to
Nat Med 1999;5:309- 13.
19
Korron DN, Ma BY, Cardoso WV et al. Bone marrow-derived cells as progenitors of lung alveolar epithelium. Development 2001;128:5181- 8.
Horwitz EM, Prockop DJ, Fitzpatrick LA et al. Transplantability and therapeutic effects of bone marrow-derived mesenchymal
18
Ono K, Takii T, Onozaki K et al. Migration of exogenous
cells repair the infarcted heart, improving function and survival.
potential source of hepatic oval cells. Science 1999;284:1168 - 70. 17
25
173
J Clin Invest
28
Williams MC, Cao Y, Hinds A et al. TI alpha protein is developmentally regulated and expressed by alveolar type I cells, choroid plexus, and ciliary epithelia of adult rats. Am
J
Respir Cell Mol BioI 1996;14:577-85. 29 Rishi AK, Joyce-Brady M, Fisher J et al. Cloning, characterization, and development expression of a rat lung alveolar type I cell gene in embryonic endodermal and neural derivatives. Dev BioI 1995;167:294- 306. 30 Danto SI, Zabski SM, Crandall ED. Reactivity of alveolar epithelial cells in primary culture with type I cell monoclonal antibodies. Am J Respir Cell Mol BioI 1992;6:296- 306. 31 Cao YX,JeanJC, Williams MC. Cytosine methylation of an Spl site contributes to organ-specific and cell-specific regulation of expression of the lung epithelial gene tlalpha. Biochem J 2000;350:883 - 90. 32 Ramirez MI, Rishi AK, Cao YX, Williams MC. TGT3, thyroid transcription factor I, and Spl elements regulate transcriptional activity of the l.3-kilobase pair promoter of TI alpha, a lung alveolar type I cell gene. J BioI Chem 1997;272:26285 - 94. 33 Costa RH, Kalinichenko VV, Lim L. Transcription factors in mouse lung development and function. Am J Physiol Lung Cell Mol Physiol 2001;280:L823-38. 34 TichelaarJW, Lim L, Costa RH, WhitserrJA. HNF-3/forkhead homologue-4 influences lung morphogenesis and respiratory epithelial cell differentiation if1 vivo. Dev BioI 1999;213:405 -17.