We have previously reported that fetal and adult skin fibroblasts display distinctive migratory phenotypes on 3-D collagen substrata and that these behavioural characteristics may be quantified by a function defined as the cell density migration index (CDMI). Subsequent work indicated that this difference in migratory phenotype was due to the production by fetal fibroblasts of a migration stimulating factor (MSF) that is not produced by normal adult skin fibroblasts. We now present data indicating that: (a) unselected fibroblasts obtained from 14/14 (100%) of adult gingival explants expressed fetal-like CDMI values compared to only 1/10 (10%) of similarly explanted paired skin cells; (b) 12/12 (100%) of these gingival fibroblast lines also produced detectable quantities of MSF compared to 0/9 (0%) of the tested skin cells; (c) by microdissection studies, gingival fibroblasts obtained from different anatomical microdomains consisted of behaviourally distinct subpopulations, with cells derived from the papillary tips (PAP fibroblasts) displaying fetal-like CDMI values and persistent MSF production, whilst cells obtained from the deeper reticular tissue (RET fibroblasts) were adult-like with respect to these two criteria; (d) PAP fibroblasts were also smaller and achieved higher saturation cell densities compared to paired RET cells; (e) PAP fibroblasts passaged in vitro underwent a fetal-to-adult phenotypic transition characterized by the adoption of various RET cell characteristics, including the acquisition of CDMI values falling within the adult range and cessation in MSF production; and (f) early passage PAP fibroblasts incubated in the presence of an affinity-purified anti-MSF rabbit polyclonal antibody were induced to alter their migratory phenotype and exhibited CDMI values falling within the adult range. Statistical analysis indicated a highly significant correlation between the expression of a fetal-like CDMI and production of MSF (P < 0.00001, using the Fisher exact contingency test). Taken together, these observations suggest that the production of MSF by PAP fibroblasts is responsible for their characteristically fetal-like migratory behaviour. The existence of such inter- and intra-site phenotypic heterogeneity in populations of skin and gingival fibroblasts is discussed in the context of fibroblast lineage relationships and the possible contribution of persistently fetal-like fibroblast subpopulations to connective tissue function in wound healing.

REFERENCES

Albini
A.
,
Pontz
B.
,
Pulz
M.
,
Allavena
G.
,
Mensing
H.
,
Muller
P. K.
(
1988
).
Decline of fibroblast chemotaxis with age of donor and cell passage number.
Coll. Rel. Res
1
,
23
37
Azzarone
B.
,
Macieira-Coehlo
A.
(
1982
).
Heterogeneity in the kinetics of proliferation within human skin fibroblast populations.
J. Cell Sci
57
,
177
187
Bartold
P. M.
,
Boyd
R. R.
,
Page
R. C.
(
1986
).
Proteoglycans synthesized by gingival fibroblasts derived from human donors of different ages.
J. Cell. Physiol
126
,
37
46
Basset
P.
,
Bellocq
J. P.
,
Wolf
C.
,
Stoll
I.
,
Hutin
P.
,
Limacher
J. M.
,
Podhajcer
O. L.
,
Chenard
M. P.
,
Rio
M. C.
,
Chambon
P.
(
1990
).
A novel metalloproteinase gene specifically expressed in stromal cells of breast carcinomas.
Nature
348
,
699
704
Bayreuther
K.
,
Rodemann
H. P.
,
Hommel
R.
,
Dittmann
K.
,
Albiez
M.
,
Francz
P. I.
(
1988
).
Human skin fibroblasts in vitro differentiate along a terminal cell lineage.
Proc. Nat. Acad. Sci. USA
85
,
5112
5116
Becker
J.
,
Schippan
D.
,
Hahn
E. G.
,
Albert
G.
,
Reichart
P.
(
1986
).
The immunohistochemical distribution of collagens type IV, V, VI and of laminin in the human oral mucosa.
Arch. Oral Biol
31
,
179
186
Bertolami
C. N.
,
Bronson
R. E.
(
1990
).
Expression of different glycosaminoglycan synthetic phenotypes by lapine dermal wound fibroblasts.
Matrix
10
,
1
9
Bordin
S.
,
Page
R. C.
,
Narayanan
A. S.
(
1984
).
Heterogeneity of normal human diploid fibroblasts: isolation and characterization of one phenotype.
Science
223
,
171
173
Brinckerhoff
C. E.
,
Nagel
J. E.
(
1981
).
Collagenase production by cloned populations of rabbit synovial fibroblasts.
Coll. Res
5
,
433
444
Bronson
R. E.
,
Argenta
J. G.
,
Siebert
E. P.
,
Bertolami
C. N.
(
1988
).
Distinctive fibroblastic subpopulations in skin oral mucosa demonstrated by difference in glycosaminoglycan content. In Vitro Cell.
Dev. Biol
24
,
1121
1126
Bronson
R. E.
,
Treat
J. A.
,
Bertolami
C. N.
(
1989
).
Fibroblastic subpopulations in uninjured and wounded rabbit oral mucosa.
J. Dent. Res
68
,
51
58
Chavrier
C.
,
Couble
M. L.
,
Magloire
H.
,
Grimaud
J. A.
(
1981
).
Immunohistochemical localisation of type I, III and IV collagen in healthy human gingiva.
J. Biol. Bucc
9
,
271
277
Chavrier
C.
,
Couble
M. L.
,
Magloire
H.
,
Grimaud
J. A.
(
1984
).
Connective tissue organisation of healthy human gingiva; ultrastructural localisation of collagen types I, III, IV.
J. Periodont. Res
19
,
221
229
Chen
J.
,
Grant
M. E.
,
Schor
A. M.
,
Schor
S. L.
(
1989
).
Differences between adult an fetal fibroblasts in the regulation of hyaluronate synthesis: correlation with migratory activity.
J. Cell Sci
94
,
577
589
Chiquet-Ehrismann
R.
,
Mackie
E. J.
,
Pearson
C. A.
,
Sakakura
T.
(
1986
).
Tenascin: An extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis.
Cell
47
,
131
139
DePalma
R. L.
,
Krummel
T. M.
,
Durham
L. A.
,
Michna
B. A.
,
Thomas
B. L.
,
Nelson
J. M.
,
Diegelmann
R. F.
(
1989
).
Characterization and quantitation of wound matrix in the fetal rabbit.
Matrix
9
,
224
231
Dvorak
H. F.
(
1986
).
Tumors: wounds that do not heal.
New Eng. J. Med
315
,
1650
1659
Ellis
I.
,
Grey
A. M.
,
Schor
A. M.
,
Schor
S. L.
(
1992
).
Antagonistic effects of transforming growth factor beta and MSF on fibroblast migration and hyaluronic acid synthesis: possible implications for dermal wound healing.
J. Cell Sci
102
,
447
456
ffrench-Constant
C.
,
Van De Water
L.
,
Dvorak
H. F.
,
Hynes
R. O.
(
1989
).
Reappearance of an embryonic pattern of fibronectin splicing during wound healing in the adult rat.
J. Cell Biol
109
,
903
914
Grey
A. M.
,
Schor
A. M.
,
Rushton
G.
,
Ellis
I.
,
Schor
S. L.
(
1989
).
Purification of the migration stimulating factor produced by fetal and breast cancer patient fibroblasts.
Proc. Nat. Acad. Sci. USA
86
,
2438
2442
Goldring
S. R.
,
Stephenson
M. L.
,
Downie
E.
,
Krane
S. M.
,
Korn
J. H.
(
1990
).
Heterogeneity in hormone response and patterns of collagen synthesis in cloned dermal fibroblasts.
J. Clin. Invest
85
,
798
803
Häkkinen
L.
,
Larjava
H.
(
1992
).
Characterization of fibroblast clones from periodontal granulation tissue in vitro.
J. Dent. Res
71
,
1901
1907
Hallock
G. G.
(
1985
).
In utero cleft lip repair in A/j mice.
Plast. Reconstr. Surg
75
,
785
788
Harper
R. A.
,
Grove
G.
(
1979
).
Human skin fibroblasts derived from papillary and reticular dermis: differences in growth potential in vitro.
Science
204
,
526
527
Harper
R. A.
(
1989
).
Keloid fibroblasts in culture: abnormal growth behaviour and altered response to epidermal growth factor.
Cell Biol. Int. Rep
13
,
325
335
Hill
D. J.
,
Strain
A. J.
,
Elstow
S. F.
,
Swenne
I.
,
Milner
R. D. G.
(
1986
).
Bi-functional action of transforming growth factor-beta on DNA synthesis in early passage human fetal fibroblasts.
J. Cell. Physiol
128
,
322
328
Hurum
S.
,
Sodek
J.
,
Aubin
J.
(
1982
).
Synthesis of collagen, collagenase and collagenase inhibitors by cloned human gingival fibroblasts and the effect of concanavalin A.
Biochem. Biophys. Res. Commun
107
,
357
366
Kischer
C. W.
,
Wagner
H. N.
,
Pindur
J.
,
Holubec
H.
, et al. 
(
1989
).
Increased fibronectin production by cell lines from hypertrophic scar and keloid.
Conn. Tiss. Res
23
,
279
288
Klein-Szanto
A.
,
Schroeder
H. E.
(
1977
).
Architecture and density of connective tissue papillae of human oral mucosa.
J. Anat
123
,
93
103
Ko
S.
,
Page
R. C.
,
Narayanan
A. S.
(
1977
).
Fibroblast heterogeneity and prostaglandin regulation of subpopulations.
Proc. Nat. Acad. Sci. USA
74
,
3429
3432
Kondo
H.
,
Kasuga
H.
,
Noumura
T.
(
1985
).
The heterogeneity of human fibroblasts as determined from the effects of hydrocortisone on cell growth and specific dexamethasone binding.
Exp. Cell Res
158
,
342
348
Korn
J. H.
(
1985
).
Substrain heterogeneity in prostaglandin E2synthesis of human dermal fibroblasts.
Arth. Rheum
28
,
315
322
Korn
J. H.
,
Brinckerhoff
C. E.
,
Edwards
C. E.
(
1985
).
Synthesis of PGE2, collagenase and tissue factor by fibroblast substrains: substrains are differentially activated for different metabolic products.
Coll. Rel. Res
5
,
437
447
Larjava
H.
,
Heino
J.
,
Kähäri
V.-M.
,
Krusius
T.
,
Vuorio
E.
(
1989
).
Characterisation of one phenotype of human periodontal granulation tissue fibroblasts.
J. Dent. Res
68
,
20
25
Lightner
V. A.
,
Slemp
C. A.
,
Erikson
H. P.
(
1990
).
Localisation and quantitation of hexabrachion (tenascin) in skin, embryonic brain, tumours and plasma.
Ann. New York Acad. Sci
580
,
260
275
Limeback
H.
,
Sodek
J.
,
Aubin
J.
(
1983
).
Variation in collagen gene expression by cloned periodontal ligament cells.
J. Periodont. Res
18
,
242
248
Mackie
E. J.
,
Halfter
W.
,
Liverani
D.
(
1988
).
Induction of tenascin in healing wounds.
J. Cell Biol
107
,
2757
2767
Martin
G. M.
,
Sprague
C. A.
,
Norwood
T. H.
,
Pendergrass
W. R.
(
1974
).
Clonal selection, attenuation and differentiation in an in vitro model of hyperplasia.
Amer. J. Pathol
74
,
137
153
Martin
M.
,
El Nabout
R.
,
Lafuma
C.
,
Crechet
F.
,
Remy
J.
(
1990
).
Fibronectin and collagen gene expression during in vitro ageing of pig skin fibroblasts.
Exp. Cell Res
191
,
8
13
Mast
B. A.
,
Haynes
J. H.
,
Krummel
T. M.
,
Diegelmann
R. F.
,
Cohen
I. K.
(
1992
).
In vivo degradation of foetal wound hyaluronic acid results in increased fibroplasia, collagen deposition, and neovascularization.
Plast. Reconstr. Surg
89
,
503
509
McCulloch
C. A. G.
,
Barghava
U.
,
Melcher
A. H.
(
1989
).
Cell death and the regulation of populations of cells in the periodontal ligament.
Cell Tiss. Res
255
,
129
138
McCulloch
C. A. G.
,
Knowles
G.
(
1991
).
Discrimination of two fibroblast progenitor populations in early explant cultures of hamster gingiva.
Cell Tiss. Res
264
,
87
94
Mollenhauer
J.
,
Bayreuther
K.
(
1986
).
Donor-age-related changes in the morphology, growth potential and collagen biosynthesis in rat fibroblast subpopulations in vitro.
Differentiation
32
,
165
172
Muggleton-Harris
A. L.
,
Reisert
P. S.
,
Burghoff
R. L.
(
1982
).
In vitro characterization of response to stimulus (wounding) with regard to ageing in human skin fibroblasts.
Mech. Ageing Dev
19
,
37
43
Nakano
S.
,
Ts'O
P. O.
(
1981
).
Cellular differentiation and neoplasia: characterization of subpopulations of cells that have neoplasia related growthproperties in Syrian hamster embryo cell cultures.
Proc. Nat. Acad. Sci. USA
78
,
4995
4999
Pascal
R. R.
,
Kaye
G. I.
,
Lane
N.
(
1968
).
Colonic pericryptal fibroblast sheath: replication, migration and cytodifferentiation of a mesenchymal cell system in adult tissue.
Gastroenterology
54
,
835
851
Pender
N.
,
Heaney
T. G.
,
Pycock
D.
,
West
C. R.
(
1988
).
Progenitor connective tissue cell populations in the gingival papilla of the rat.
J. Periodont. Res
23
,
175
181
Pienta
K.
,
Coffey
D. S.
(
1990
).
Characterization of the subtypes of cell motility in ageing human skin fibroblasts.
Mech. Ageing Dev
56
,
99
105
Picardo
M.
,
Schor
S. L.
,
Grey
A.-M.
,
Howell
A.
,
Laidlaw
I.
,
Redford
J.
,
Schor
A. M.
(
1991
).
Migration stimulating activity in serum of breast cancer patients.
Lancet
337
,
130
133
Picardo
M.
,
McGurk
M.
,
Schor
S. L.
,
Grey
A.-M.
,
Ellis
I.
) (
1992
).
Identification of migration stimulating factor in wound fluid.
Exp. Mol. Path
57
,
8
21
Russell
S. B.
,
Trupin
K. M.
,
Rodriguez-Eaton
S.
,
Russell
J. D.
,
Trupin
J. S.
(
1988
).
Reduced growth factor requirement of keloid-derived fibroblasts may account for tumour growth.
Proc Nat. Acad. Sci. USA
85
,
587
591
Schafer
I. A.
,
Pandy
M.
,
Ferguson
R.
,
Davis
B. R.
(
1985
).
Comparative observation on fibroblasts derived from the papillary and reticular dermis of infants and adults; growth kinetics, packing density at confluence and surface morphology.
Mech. Ageing Dev
33
,
115
137
Schafer
I. A.
,
Shapiro
A.
,
Kovach
M.
,
Lang
C.
,
Fratianne
R. B.
(
1989
).
The interaction of human papillary and reticular fibroblasts and human keratinocytes in the contraction of three-dimensional floating collagen lattices.
Exp. Cell Res
183
,
112
125
Schor
S. L.
(
1980
).
Cell proliferation and migration within three-dimensional collagen gels.
J. Cell Sci
41
,
159
175
Schor
S. L.
,
Schor
A. M.
,
Rushton
G.
,
Smith
L.
(
1985
).
Adult, foetal, and transformed fibroblasts display different migratory phenotypes on collagen gels: Evidence for an isoformic transition during foetal development.
J. Cell Sci
73
,
221
234
Schor
S. L.
,
Schor
A. M.
,
Durning
P.
,
Rushton
G.
(
1985
).
Skin fibroblasts obtained from cancer patients display foetal-like migratory behaviour on collagen gels.
J. Cell Sci
73
,
235
244
Schor
S. L.
,
Haggie
J.
,
Durning
P.
,
Howell
A.
,
Sellwood
R. A. S.
,
Crowther
D.
(
1986
).
The occurrence of a foetal fibroblast phenotype in breast cancer.
Int. J. Cancer
37
,
831
836
Schor
S. L.
,
Schor
A. M.
(
1987
).
Clonal heterogeneity in fibroblasts: implications for the control of epithelial-mesenchymal interactions.
BioEssays
7
,
200
204
Schor
S. L.
,
Schor
A. M.
,
Howell
A.
,
Crowther
D.
(
1987
).
Hypothesis: persistent expression of fetal phenotypic characteristics by fibroblasts is associated with an increased susceptibility to neoplastic disease.
Exp. Cell Biol
55
,
11
17
Schor
S. L.
,
Schor
A. M.
,
Grey
A. M.
,
Rushton
G.
(
1988
).
Fetal and cancer patient fibroblasts produce an autocrine migration stimulating factor not made by normal adult cells.
J. Cell Sci
90
,
391
99
Schor
S. L.
,
Schor
A. M.
,
Rushton
G.
(
1988
).
Fibroblasts from cancer patients display a mixture of both foetal and adult-like phenotypic characteristics.
J. Cell Sci
90
,
401
407
Sloan
P.
,
Schor
S. L.
,
Lopes
V.
,
Chiquet-Ehrismann
R.
(
1989
).
Immunohistochemical study of the heterogeneity of tenascin distribution within the oral mucosa of the mouse.
Arch. Oral Biol
35
,
67
70
Sottile
J.
,
Mann
D. M.
,
Diemar
V.
,
Millis
A. J. T.
(
1989
).
Regulation of collagenase and collagenase mRNA production by early-and late-passage human diploid fibroblasts.
J. Cell. Physiol
138
,
281
290
Smith
J. R.
,
Pereira-Smith
O. M.
,
Schneider
E. L.
(
1978
).
Colony size distributions as a measure of in vivo and in vitro ageing.
Proc. Nat. Acad. Sci. USA
75
,
1353
1356
Stoker
M.
,
Gherardi
E.
(
1991
).
Regulation of cell movement: the motogenic cytokines.
Biochim. Biophys. Acta
1072
,
81
102
Tajima
S.
,
Pinnell
S. R.
(
1981
).
Collagen synthesis by human skin fibroblasts explanted from papillary and reticular dermis.
J. Invest. Dermatol
77
,
410
412
Thompson
R. G.
,
Nickel
B.
,
Finlayson
S.
,
Meuser
R.
,
Hamerton
J. L.
,
Wrogemann
K.
(
1983
).
56K fibroblast protein not specific for Duchenne muscular dystrophy but for skin biopsy site.
Nature
304
,
740
741
Vande Berg
J. S.
,
Rudolph
R.
,
Poolman
W. L.
,
Dsharoon
D. R.
(
1989
).
Comparative growth dynamics and actin concentration between cultured human myofibroblasts from granulating wounds and dermal fibroblasts from normal skin.
Lab. Invest
61
,
532
538
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