The therapeutic potential of ex vivo expanded CD133+ cells derived from human peripheral blood for peripheral nerve injuries

Laboratory investigation

Shin Ohtsubo M.D. 1 , Masakazu Ishikawa M.D., Ph.D. 1 , Naosuke Kamei M.D., Ph.D. 1 , 2 , Yasumu Kijima M.D., Ph.D. 3 , Osami Suzuki M.D., Ph.D. 1 , Toru Sunagawa M.D., Ph.D. 4 , Yukihito Higashi M.D., Ph.D. 2 , Haruchika Masuda M.D., Ph.D. 5 , Takayuki Asahara M.D., Ph.D. 5 , 6 and Mitsuo Ochi M.D., Ph.D. 1
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  • 1 Departments of Orthopaedic Surgery and
  • 4 Anatomy and Histology, Graduate School of Biomedical Sciences, Hiroshima University;
  • 2 Division of Regeneration and Medicine, Hiroshima University Hospital;
  • 3 Department of Orthopaedic Surgery, Sera Central Hospital, Hiroshima;
  • 5 Regenerative Medicine Science, Tokai University School of Medicine, Isehara; and
  • 6 Group of Vascular Regeneration, Institute of Biomedical Research and Innovation, Kobe, Japan
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Object

CD133+ cells have the potential to enhance histological and functional recovery from peripheral nerve injury. However, the number of CD133+ cells safely obtained from human peripheral blood is extremely limited. To address this issue, the authors expanded CD133+ cells derived from human peripheral blood using the serum-free expansion culture method and transplanted these ex vivo expanded cells into a model of sciatic nerve defect in rats. The purpose of this study was to determine the potential of ex vivo expanded CD133+ cells to induce or enhance the repair of injured peripheral nerves.

Methods

Phosphate-buffered saline (PBS group [Group 1]), 105 fresh CD133+ cells (fresh group [Group 2]), 105 ex vivo expanded CD133+ cells (expansion group [Group 3]), or 104 fresh CD133+ cells (low-dose group [Group 4]) embedded in atelocollagen gel were transplanted into a silicone tube that was then used to bridge a 15-mm defect in the sciatic nerve of athymic rats (10 animals per group). At 8 weeks postsurgery, histological and functional evaluations of the regenerated tissues were performed.

Results

After 1 week of expansion culture, the number of cells increased 9.6 ± 3.3–fold. Based on the fluorescence-activated cell sorting analysis, it was demonstrated that the initial freshly isolated CD133+ cell population contained 93.22% ± 0.30% CD133+ cells and further confirmed that the expanded cells had a purity of 59.02% ± 1.58% CD133+ cells. However, the histologically and functionally regenerated nerves bridging the defects were recognized in all rats in Groups 2 and 3 and in 6 of 10 rats in Group 4. The nerves did not regenerate to bridge the defect in any of the rats in Group 1.

Conclusions

The authors' results show that ex vivo expanded CD133+ cells derived from human peripheral blood have a therapeutic potential similar to fresh CD133+ cells for peripheral nerve injuries. The ex vivo procedure that can be used to expand CD133+ cells without reducing their function represents a novel method for developing cell therapy for nerve defects in a clinical setting.

Abbreviations used in this paper:CMAP = compound muscle action potential; FACS = fluorescence-activated cell sorting; PBS = phosphate-buffered saline.

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Contributor Notes

Address correspondence to: Shin Ohtsubo, M.D., Department of Orthopaedic Surgery, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8551, Japan. email: ohtsubo0529@yahoo.co.jp.

Please include this information when citing this paper: published online August 10, 2012; DOI: 10.3171/2012.7.JNS111503.

  • 1

    Asahara T, , Murohara T, , Sullivan A, , Silver M, , van der Zee R, & Li T, : Isolation of putative progenitor endothelial cells for angiogenesis. Science 275:964967, 1997

    • Search Google Scholar
    • Export Citation
  • 2

    Bryan DJ, , Wang KK, & Chakalis-Haley DP: Effect of Schwann cells in the enhancement of peripheral-nerve regeneration. J Reconstr Microsurg 12:4396, 1996

    • Search Google Scholar
    • Export Citation
  • 3

    Deshpande DM, , Kim YS, , Martinez T, , Carmen J, , Dike S, & Shats I, : Recovery from paralysis in adult rats using embryonic stem cells. Ann Neurol 60:3244, 2006

    • Search Google Scholar
    • Export Citation
  • 4

    Goussetis E, , Theodosaki M, , Paterakis G, , Peristeri J, , Petropoulos D, & Kitra V, : A functional hierarchy among the CD34+ hematopoietic cells based on in vitro proliferative and differentiative potential of AC133+CD34(bright) and AC133(dim/)-CD34+ human cord blood cells. J Hematother Stem Cell Res 9:827840, 2000

    • Search Google Scholar
    • Export Citation
  • 5

    Hadlock TA, , Sundback CA, , Hunter DA, , Vacanti JP, & Cheney ML: A new artificial nerve graft containing rolled Schwann cell monolayers. Microsurgery 21:96101, 2001

    • Search Google Scholar
    • Export Citation
  • 6

    Heiss C, , Keymel S, , Niesler U, , Ziemann J, , Kelm M, & Kalka C: Impaired progenitor cell activity in age-related endothelial dysfunction. J Am Coll Cardiol 45:14411448, 2005

    • Search Google Scholar
    • Export Citation
  • 7

    Iwasaki H, , Kawamoto A, , Ishikawa M, , Oyamada A, , Nakamori S, & Nishimura H, : Dose-dependent contribution of CD34-positive cell transplantation to concurrent vasculogenesis and cardiomyogenesis for functional regenerative recovery after myocardial infarction. Circulation 113:13111325, 2006

    • Search Google Scholar
    • Export Citation
  • 8

    Kawamoto A, , Katayama M, , Handa N, , Kinoshita M, , Takano H, & Horii M, : Intramuscular transplantation of G-CSFmobilized CD34+ cells in patients with critical limb ischemia: a phase I/IIa, multicenter, single-blinded, dose-escalation clinical trial. Stem Cells 27:28572864, 2009

    • Search Google Scholar
    • Export Citation
  • 9

    Kijima Y, , Ishikawa M, , Sunagawa T, , Nakanishi K, , Kamei N, & Yamada K, : Regeneration of peripheral nerve after transplantation of CD133+ cells derived from human peripheral blood. Laboratory investigation. J Neurosurg 110:758767, 2009

    • Search Google Scholar
    • Export Citation
  • 10

    Loges S, , Fehse B, , Brockmann MA, , Lamszus K, , Butzal M, & Guckenbiehl M, : Identification of the adult human hemangioblast. Stem Cells Dev 13:229242, 2004

    • Search Google Scholar
    • Export Citation
  • 11

    Mackinnon SE, & Dellon AL: Clinical nerve reconstruction with a bioabsorbable polyglycolic acid tube. Plast Reconstr Surg 85:419424, 1990

    • Search Google Scholar
    • Export Citation
  • 12

    Majka M, , Janowska-Wieczorek A, , Ratajczak J, , Ehrenman K, , Pietrzkowski Z, & Kowalska MA, : Numerous growth factors, cytokines, and chemokines are secreted by human CD34+ cells, myeloblasts, erythroblasts, and megakaryoblasts and regulate normal hematopoiesis in an autocrine/paracrine manner. Blood 97:30753085, 2001

    • Search Google Scholar
    • Export Citation
  • 13

    Masuda H, , Iwasaki H, , Kawamoto A, , Akimaru H, , Ishikawa M, & Asahara T, : Development of serum-free quality and quantity control culture of colony forming endothelial progenitor cell expansion for vasculogenesis. Stem Cells Trans Med [epub ahead of print] 2012

    • Search Google Scholar
    • Export Citation
  • 14

    Matsumoto T, , Kawamoto A, , Kuroda R, , Ishikawa M, , Mifune Y, & Iwasaki H, : Therapeutic potential of vasculogenesis and osteogenesis promoted by peripheral blood CD34-positive cells for functional bone healing. Am J Pathol 169:14401457, 2006

    • Search Google Scholar
    • Export Citation
  • 15

    Murakami T, , Fujimoto Y, , Yasunaga Y, , Ishida O, , Tanaka N, & Ikuta Y, : Transplanted neuronal progenitor cells in a peripheral nerve gap promote nerve repair. Brain Res 974:1724, 2003

    • Search Google Scholar
    • Export Citation
  • 16

    Peichev M, , Naiyer AJ, , Pereira D, , Zhu Z, , Lane WJ, & Rafii S, : Expression of VEGFR-2 and AC133 by circulating human CD34+ cells identifies a population of functional endothelial precursors. Blood 95:952958, 2000

    • Search Google Scholar
    • Export Citation
  • 17

    Rafii S, & Lyden D: Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat Med 9:702712, 2003

    • Search Google Scholar
    • Export Citation
  • 18

    Tohill M, , Mantovani C, , Wiberg M, & Terenghi G: Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration. Neurosci Lett 362:200203, 2004

    • Search Google Scholar
    • Export Citation
  • 19

    Urbich C, & Dimmeler S: Endothelial progenitor cells functional characterization. Trends Cardiovasc Med 14:318322, 2004

  • 20

    Vasa M, , Fichtlscherer S, , Aicher A, , Adler K, , Urbich C, & Martin H, : Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease. Circ Res 89:E1E7, 2001

    • Search Google Scholar
    • Export Citation
  • 21

    Xu Y, , Tamamaki N, , Noda T, , Kimura K, , Itokazu Y, & Matsumoto N, : Neurogenesis in the ependymal layer of the adult rat 3rd ventricle. Exp Neurol 192:251264, 2005

    • Search Google Scholar
    • Export Citation
  • 22

    Yang LY, , Zheng JK, , Wang CY, & Li WY: Differentiation of adult human bone marrow mesenchymal stem cells into Schwannlike cells in vitro. Chin J Traumatol 8:7780, 2005

    • Search Google Scholar
    • Export Citation
  • 23

    Yin AH, , Miraglia S, , Zanjani ED, , Almeida-Porada G, , Ogawa M, & Leary AG, : AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 90:50025012, 1997

    • Search Google Scholar
    • Export Citation

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