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Cell Surface Expression of Stem Cell Antigen-1 (Sca-1) Distinguishes Osteo-, Chondro-, and Adipoprogenitors in Fetal Mouse Calvaria

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Abstract

The flat bones of the skull (calvaria) develop by balanced cell proliferation and differentiation in the calvarial sutures and the bone tips. As the brain grows and the calvaria expand, cells within the sutures must remain undifferentiated to maintain suture patency, but osteoprogenitors also need to be recruited into the osteogenic fronts. The exact identity of calvarial osteoprogenitors is currently not known. We used immunomagnetic cell sorting to isolate Sca-1+ and Sca-1− cells from fetal mouse calvaria and determined their differentiation potential in in vitro differentiation asssays and in vivo subcutaneous transplantations. Cells within the Sca-1+ cell fraction have a higher adipogenic potential, whereas cells within the Sca-1− cell fraction have a higher osteogenic and chondrogenic potential. The Sca-1− fraction retains its chondrogenic potential after in vitro expansion but not its osteogenic potential. The Sca-1+ fraction does not retain its adipogenic potential after in vitro expansion. Subcutaneous transplantation resulted in islands of bone and cartilage in implants that had been seeded with Sca-1− cells. In conclusion, immunomagnetic cell sorting with Sca-1 antibodies can be used to separate a Sca-1+ cell fraction with adipogenic potential from a Sca-1− cell fraction with osteogenic and chondrogenic potential. Isolation of pure populations of calvarial adipoprogenitors, osteoprogenitors, and chondroprogenitors will be beneficial for cellular studies of calvarial development, adipogenesis, osteogenesis, and chondrogenesis. Calvaria-derived osteogenic cell populations may be useful in craniofacial tissue regeneration and repair.

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References

  1. Opperman LA (2000) Cranial sutures as intramembranous bone growth sites. Dev Dyn 219:472–485

    Article  PubMed  CAS  Google Scholar 

  2. Aguila HL, Weissman IL (1996) Hematopoietic stem cells are not direct cytotoxic targets of natural killer cells. Blood 87:1225–1231

    PubMed  CAS  Google Scholar 

  3. Bachar-Lustig E, Li HW, Gur H, Krauthgamer R, Marcus H, Reisner Y (1999) Induction of donor-type chimerism and transplantation tolerance across major histocompatibility barriers in sublethally irradiated mice by Sca-1+Lin− bone marrow progenitor cells: synergism with non-alloreactive (host x donor)F1 T cells. Blood 94:3212–3221

    PubMed  CAS  Google Scholar 

  4. Bailey AS, Jiang S, Afentoulis M, Baumann CI, Schroeder DA, Olson SB, Wong MH, Fleming WH (2004) Transplanted adult hematopoietic stems cells differentiate into functional endothelial cells. Blood 103:13–19

    Article  PubMed  CAS  Google Scholar 

  5. Gussoni E, Soneoka Y, Strickland CD, Buzney EA, Khan MK, Flint AF, Kunkel LM, Mulligan RC (1999) Dystrophin expression in the mdx mouse restored by stem cell transplantation. Nature 401:390–394

    PubMed  CAS  Google Scholar 

  6. Lagasse E, Connors H, Al-Dhalimy M, Reitsma M, Dohse M, Osborne L, Wang X, Finegold M, Weissman IL, Grompe M (2000) Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med 6:1229–1234

    Article  PubMed  CAS  Google Scholar 

  7. Spangrude GJ, Heimfeld S, Weissman IL (1988) Purification and characterization of mouse hematopoietic stem cells. Science 241:58–62

    Article  PubMed  CAS  Google Scholar 

  8. Welm BE, Tepera SB, Venezia T, Graubert TA, Rosen JM, Goodell MA (2002) Sca-1(pos) cells in the mouse mammary gland represent an enriched progenitor cell population. Dev Biol 245:42–56

    Article  PubMed  CAS  Google Scholar 

  9. Baddoo M, Hill K, Wilkinson R, Gaupp D, Hughes C, Kopen GC, Phinney DG (2003) Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection. J Cell Biochem 89:1235–1249

    Article  PubMed  CAS  Google Scholar 

  10. Gojo S, Gojo N, Takeda Y, Mori T, Abe H, Kyo S, Hata J, Umezawa A (2003) In vivo cardiovasculogenesis by direct injection of isolated adult mesenchymal stem cells. Exp Cell Res 288:51–59

    Article  PubMed  CAS  Google Scholar 

  11. Lee J, Kuroda S, Shichinohe H, Ikeda J, Seki T, Hida K, Tada M, Sawada K, Iwasaki Y (2003) Migration and differentiation of nuclear fluorescence-labeled bone marrow stromal cells after transplantation into cerebral infarct and spinal cord injury in mice. Neuropathology 23:169–180

    Article  PubMed  Google Scholar 

  12. Peister A, Mellad JA, Larson BL, Hall BM, Gibson LF, Prockop DJ (2004) Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood 103:1662–1668

    Article  PubMed  CAS  Google Scholar 

  13. Park J, Gelse K, Frank S, von der Mark K, Aigner T, Schneider H (2006) Transgene-activated mesenchymal cells for articular cartilage repair: a comparison of primary bone marrow-, perichondrium/periosteum- and fat-derived cells. J Gene Med 8:112–125

    Article  PubMed  CAS  Google Scholar 

  14. Wolnicka-Glubisz A, King W, Noonan FP (2005) SCA-1+ cells with an adipocyte phenotype in neonatal mouse skin. J Invest Dermatol 125:383–385

    Article  PubMed  CAS  Google Scholar 

  15. Van Vlasselaer P, Falla N, Snoeck H, Mathieu E (1994) Characterization and purification of osteogenic cells from murine bone marrow by two-color cell sorting using anti-Sca-1 monoclonal antibody and wheat germ agglutinin. Blood 84:753–763

    PubMed  Google Scholar 

  16. Horowitz MC, Fields A, DeMeo D, Qian HY, Bothwell AL, Trepman E (1994) Expression and regulation of Ly-6 differentiation antigens by murine osteoblasts. Endocrinology 135:1032–1043

    Article  PubMed  CAS  Google Scholar 

  17. Ito CY, Li CY, Bernstein A, Dick JE, Stanford WL (2003) Hematopoietic stem cell and progenitor defects in Sca-1/Ly-6A-null mice. Blood 101:517–523

    Article  PubMed  CAS  Google Scholar 

  18. Bonyadi M, Waldman SD, Liu D, Aubin JE, Grynpas MD, Stanford WL (2003) Mesenchymal progenitor self-renewal deficiency leads to age-dependent osteoporosis in Sca-1/Ly-6A null mice. Proc Natl Acad Sci USA 100:5840–5845

    Article  PubMed  CAS  Google Scholar 

  19. Mitchell PO, Mills T, O’Connor RS, Graubert T, Dzierzak E, Pavlath GK (2005) Sca-1 negatively regulates proliferation and differentiation of muscle cells. Dev Biol 283:240–252

    Article  PubMed  CAS  Google Scholar 

  20. Epting CL, Lopez JE, Shen X, Liu L, Bristow J, Bernstein HS (2004) Stem cell antigen-1 is necessary for cell-cycle withdrawal and myoblast differentiation in C2C12 cells. J Cell Sci 117:6185–6195

    Article  PubMed  CAS  Google Scholar 

  21. Bianco P, Riminucci M, Gronthos S, Robey PG (2001) Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells 19:180–192

    Article  PubMed  CAS  Google Scholar 

  22. Krebsbach PH, Kuznetsov SA, Bianco P, Robey PG (1999) Bone marrow stromal cells: characterization and clinical application. Crit Rev Oral Biol Med 10:165–181

    Article  PubMed  CAS  Google Scholar 

  23. Long MW (2001) Osteogenesis and bone-marrow-derived cells. Blood Cells Mol Dis 27:677–690

    Article  PubMed  CAS  Google Scholar 

  24. Pettway GJ, Schneider A, Koh AJ, Widjaja E, Morris MD, Meganck JA, Goldstein SA, McCauley LK (2005) Anabolic actions of PTH (1–34): use of a novel tissue engineering model to investigate temporal effects on bone. Bone 36:959–970

    Article  PubMed  CAS  Google Scholar 

  25. Rosen ED, Spiegelman BM (2000) Molecular regulation of adipogenesis. Annu Rev Cell Dev Biol 16:145–171

    Article  PubMed  CAS  Google Scholar 

  26. Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T (1997) Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 89:755–764

    Article  PubMed  CAS  Google Scholar 

  27. Stein GS, Lian JB, van Wijnen AJ, Stein JL, Montecino M, Javed A, Zaidi SK, Young DW, Choi JY, Pockwinse SM (2004) Runx2 control of organization, assembly and activity of the regulatory machinery for skeletal gene expression. Oncogene 23:4315–4329

    Article  PubMed  CAS  Google Scholar 

  28. Steenhuis P, Carr KM, Pettway GJ, Ignelzi MA Stem/Progenitor Cells Isolated From Postnatal Mouse Calvaria. Unpublished results

  29. Wang J, Glimcher MJ (1999) Characterization of matrix-induced osteogenesis in rat calvarial bone defects: II. Origins of bone-forming cells. Calcif Tissue Int 65:486–493

    Article  PubMed  CAS  Google Scholar 

  30. Opperman LA, Passarelli RW, Morgan EP, Reintjes M, Ogle RC (1995) Cranial sutures require tissue interactions with dura mater to resist osseous obliteration in vitro. J Bone Miner Res 10:1978–1987

    Article  PubMed  CAS  Google Scholar 

  31. Opperman LA, Sweeney TM, Redmon J, Persing JA, Ogle RC (1993) Tissue interactions with underlying dura mater inhibit osseous obliteration of developing cranial sutures. Dev Dyn 198:312–322

    PubMed  CAS  Google Scholar 

  32. Mabbutt LW, Kokich VG (1979) Calvarial and sutural re-development following craniectomy in the neonatal rabbit. J Anat 129:413–422

    PubMed  CAS  Google Scholar 

  33. Mabbutt LW, Kokich VG, Moffett BC, Loeser JD (1979) Subtotal neonatal calvariectomy. A radiographic and histological evaluation of calvarial and sutural redevelopment in rabbits. J Neurosurg 51:691–696

    PubMed  CAS  Google Scholar 

  34. Hobar PC, Schreiber JS, McCarthy JG, Thomas PA (1993) The role of the dura in cranial bone regeneration in the immature animal. Plast Reconstr Surg 92:405–410

    Article  PubMed  CAS  Google Scholar 

  35. Drake DB, Persing JA, Berman DE, Ogle RC (1993) Calvarial deformity regeneration following subtotal craniectomy for craniosynostosis: a case report and theoretical implications. J Craniofac Surg 4:85–90

    Article  PubMed  CAS  Google Scholar 

  36. Hutmacher DW, Sittinger M (2003) Periosteal cells in bone tissue engineering. Tissue Eng 9 Suppl 1:S45–S64

    Article  PubMed  Google Scholar 

  37. Nakahara H, Bruder SP, Goldberg VM, Caplan AI (1990) In vivo osteochondrogenic potential of cultured cells derived from the periosteum. Clin Orthop Relat Res 259:223–232

    PubMed  Google Scholar 

  38. Aubin JE (1999) Osteoprogenitor cell frequency in rat bone marrow stromal populations: role for heterotypic cell–cell interactions in osteoblast differentiation. J Cell Biochem 72:396–410

    Article  PubMed  CAS  Google Scholar 

  39. Purpura KA, Zandstra PW, Aubin JE (2003) Fluorescence activated cell sorting reveals heterogeneous and cell non-autonomous osteoprogenitor differentiation in fetal rat calvaria cell populations. J Cell Biochem 90:109–120

    Article  PubMed  CAS  Google Scholar 

  40. Eipers PG, Kale S, Taichman RS, Pipia GG, Swords NA, Mann KG, Long MW (2000) Bone marrow accessory cells regulate human bone precursor cell development. Exp Hematol 28:815–825

    Article  PubMed  CAS  Google Scholar 

  41. Stewart K, Monk P, Walsh S, Jefferiss CM, Letchford J, Beresford JN (2003) STRO-1, HOP-26 (CD63), CD49a and SB-10 (CD166) as markers of primitive human marrow stromal cells and their more differentiated progeny: a comparative investigation in vitro. Cell Tissue Res 313:281–290

    Article  PubMed  CAS  Google Scholar 

  42. Goodell MA, McKinney-Freeman S, Camargo FD (2004) Isolation and characterization of side population cells. Methods Mol Biol 290:343–352

    Google Scholar 

  43. Smith JD, Abramson M (1974) Membranous vs. endochondral bone autografts. Arch Otolaryngol 99:203–205

    PubMed  CAS  Google Scholar 

  44. Zins JE, Whitaker LA (1983) Membranous versus endochondral bone: implications for craniofacial reconstruction. Plast Reconstr Surg 72:778–785

    Article  PubMed  CAS  Google Scholar 

  45. Tessier P (1982) Autogenous bone grafts taken from the calvarium for facial and cranial applications. Clin Plast Surg 9:531–538

    PubMed  CAS  Google Scholar 

  46. Hunter D, Baker S, Sobol SM (1990) Split calvarial grafts in maxillofacial reconstruction. Otolaryngol Head Neck Surg 102:345–350

    PubMed  CAS  Google Scholar 

  47. Frodel JL Jr, Marentette LJ, Quatela VC, Weinstein GS (1993) Calvarial bone graft harvest. Techniques, considerations, and morbidity. Arch Otolaryngol Head Neck Surg 119:17–23

    PubMed  Google Scholar 

  48. Carinci F, Farina A, Zanetti U, Vinci R, Negrini S, Calura G, Laino G, Piattelli A (2005) Alveolar ridge augmentation: a comparative longitudinal study between calvaria and iliac crest bone grafrs. J Oral Implantol 31:39–45

    Article  PubMed  Google Scholar 

  49. Cinberg JZ, Rosenbaum FA, Lowrie C, Gorman M (1985) Calvarial grafts for midface rehabilitation. Arch Otolaryngol 111:434–436

    PubMed  CAS  Google Scholar 

  50. Maves MD, Matt BH (1986) Calvarial bone grafting of facial defects. Otolaryngol Head Neck Surg 95:464–470

    PubMed  CAS  Google Scholar 

  51. Smolka W, Eggensperger N, Kollar A, Iizuka T (2005) Midfacial reconstruction using calvarial split bone grafts. Arch Otolaryngol Head Neck Surg 131:131–136

    Article  PubMed  Google Scholar 

  52. Frodel JL (1999) Calvarial bone graft harvest in children. Otolaryngol Head Neck Surg 121:78–81

    Article  PubMed  CAS  Google Scholar 

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Acknowledgement

We thank Laurie K. McCauley and Michael W. Long for valuable discussions and Kathy Welch for advice on statistical methods. This study was supported by the National Institutes of Health (DE11530).

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Authors and Affiliations

  1. Department of Orthodontics and Pediatric Dentistry, University of Michigan School of Dentistry, 1011 North University Avenue, Ann Arbor, MI, 48109-1078, USA

    Pieter Steenhuis & Michael A. Ignelzi Jr.

  2. Department of Periodontics and Oral Medicine, School of Dentistry, and Department of Biomedical Engineering, College of Engineering, University of Michigan, 1011 North University Avenue, Ann Arbor, MI, 48109-1078, USA

    Glenda J. Pettway

  3. Department of Dermatology, University of California Davis, 1 Shields Avenue, TB205, Room 102, Davis, CA, 95616-5270, USA

    Pieter Steenhuis

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  1. Pieter Steenhuis
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Correspondence to Pieter Steenhuis.

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Steenhuis, P., Pettway, G.J. & Ignelzi, M.A. Cell Surface Expression of Stem Cell Antigen-1 (Sca-1) Distinguishes Osteo-, Chondro-, and Adipoprogenitors in Fetal Mouse Calvaria. Calcif Tissue Int 82, 44–56 (2008). https://doi.org/10.1007/s00223-007-9083-4

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