HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Askenasy, N. Articles by Farkas, D. L. Search for Related Content PubMed PubMed Citation Articles by Askenasy, N. Articles by Farkas, D. L.

Optical Imaging of PKH-Labeled Hematopoietic Cells in Recipient Bone Marrow In Vivo

^a Frankel Laboratory for Bone Marrow Transplantation, Center for Stem Cell Research, Schneider Children’s Medical Center of Israel, Petach Tikva, Israel;
^b McGowan Institute for Regenerative Medicine and
^c Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania, USA

Key Words. In vivo tracking • Fluorescence microscopy • Laser tweezers • Energy transfer • Bone marrow transplantation • PKH membrane linkers

Nadir Askenasy, M.D., Department of Pediatric Hematology Oncology, Schneider Children’s Medical Center of Israel, 14 Kaplan Street, Petach Tikva 49202 Israel. Telephone: 972-3-641-1475; Fax: 972-3-641-1475; e-mail: anadir{at}012.net.il

This work describes an optical technique for characterization of the early stages of hematopoietic stem cell (HSC) engraftment under physiological conditions and in real time. Bone marrow cells (BMCs) labeled with PKH membrane linkers were injected into conditioned recipients (B10B10.BR mice) preoperated for placement of optical windows over femoral epiphyses. Labeled cells were tracked in vivo by fluorescence microscopy. Cellular adhesion to the BM stroma was tested with laser tweezers, and viability was assayed by the propidium iodide (PI) exclusion test, as determined from energy-transfer measurements of the pair PKH67-PI in freshly excised femurs in situ. At optimal concentrations for in vivo tracking, 1-4 µM PKH dyes neither impaired the viability of BMCs nor the capacity of allogeneic HSCs to reconstitute hematopoiesis in myeloablated recipients. The optical window allowed in vivo visualization of 23%-26% of the PKH-labeled BMCs in the femur. The homing efficiencies at 16 hours posttransplantation were quantified as 1.77% ± 0.15% and 0.21% ± 0.02% for syngeneic and allogeneic BMCs, respectively. In femurs excised 16 hours after transplantation, 70% ± 9% of the cells were adherent to the BM stroma, and two-thirds of the cells were PI negative (viable). In vivo tracking and in situ assessment of labeled HSCs in recipient BM provide important quantitative and qualitative insights into the early stages of engraftment. Correlation of early events and the efficiency of durable engraftment serve as the basis for a systematic approach toward optimization of the conditions for transplantation.

This article has been cited by other articles:

				Y. Hayashi, S. Tsuji, M. Tsujii, T. Nishida, S. Ishii, H. Iijima, T. Nakamura, H. Eguchi, E. Miyoshi, N. Hayashi, et al. Topical Implantation of Mesenchymal Stem Cells Has Beneficial Effects on Healing of Experimental Colitis in Rats J. Pharmacol. Exp. Ther., August 1, 2008; 326(2): 523 - 531. [Abstract] [Full Text] [PDF]

				Y. Hayashi, S. Tsuji, M. Tsujii, T. Nishida, S. Ishii, H. Iijima, T. Nakamura, H. Eguchi, E. Miyoshi, N. Hayashi, et al. Topical transplantation of mesenchymal stem cells accelerates gastric ulcer healing in rats Am J Physiol Gastrointest Liver Physiol, March 1, 2008; 294(3): G778 - G786. [Abstract] [Full Text] [PDF]

				D. N. Haylock, B. Williams, H. M. Johnston, M. C.P. Liu, K. E. Rutherford, G. A. Whitty, P. J. Simmons, I. Bertoncello, and S. K. Nilsson Hemopoietic Stem Cells with Higher Hemopoietic Potential Reside at the Bone Marrow Endosteum Stem Cells, April 1, 2007; 25(4): 1062 - 1069. [Abstract] [Full Text] [PDF]

				R. Zhou, P. D. Acton, and V. A. Ferrari Imaging Stem Cells Implanted in Infarcted Myocardium J. Am. Coll. Cardiol., November 21, 2006; 48(10): 2094 - 2106. [Abstract] [Full Text] [PDF]

				T. Kanamori, G. Watanabe, T. Yasuda, H. Nagamine, H. Kamiya, and Y. Koshida Hybrid Surgical Angiogenesis: Omentopexy Can Enhance Myocardial Angiogenesis Induced by Cell Therapy Ann. Thorac. Surg., January 1, 2006; 81(1): 160 - 167. [Abstract] [Full Text] [PDF]

				T. Lapidot, A. Dar, and O. Kollet How do stem cells find their way home? Blood, September 15, 2005; 106(6): 1901 - 1910. [Abstract] [Full Text] [PDF]

				J. V. Frangioni and R. J. Hajjar In Vivo Tracking of Stem Cells for Clinical Trials in Cardiovascular Disease Circulation, November 23, 2004; 110(21): 3378 - 3383. [Abstract] [Full Text] [PDF]