EXPRESSIONS OF B-CELL-SPECIFIC MONOCLONAL LEUKEMIA VIRUS INSERT SITE 1 AND HUMAN TELOMERASE REVERSE TRANSCRIPTASE GENES IN CD34+ CELLS DURING EX VIVO EXPANSION PROCESS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

GE Jianyun , CAI Haibo , DU Zheng , TAN Wensong.

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, P.R.China. Corresponding author: CAI Haibo, E-mail: caihaibo@ecust.edu. cn;

Corresponding author：

Keywords：

CD34+ cell Ex vivo expansion; B-cell-specific monoclonal leukemia virus insert site 1; Human telomerase reverse transcriptase

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ObjectiveTo investigate the relationship between the expressions of B-cell-specific monoclonal leukemia virus insert site 1 (Bmi1) and human telomerase reverse transcriptase (hTERT) genes and the proliferative capacity of stem cells. MethodsCord blood CD34+ cells were cultured in IMDM medium containing fetal bovine serum, stem cell growth factor, thrombopoietin, and Fms-like tyrosine kinase 3 ligand during a 28-day ex vivo expansion process, while the expansion fold, specific growth rate, and the colony-forming efficiency were monitored. Then the Bmi1 and hTERT mRNA expressions in CD34+ cells were detected by fluorescence quantitative PCR, and the relations between the expressions and the cell proliferative capacity were analyzed. ResultsCD34+ cells expanded (20.1 ± 3.5) folds during the 28-day culture, while the proportion declined from 95.5% ± 2.6% before culture to 2.1% ± 0.4%. Both the specific growth rate and colony-forming efficiency of CD34+ cells declined significantly after 7 days, while the expression levels of Bmi1 and hTERT mRNA in CD34+ cells reached top at 7 days and declined gradually thereafter. ConclusionThe expressions of Bmi1 and hTERT genes may have a close relation to the proliferative capacity of CD34+ cells.

Citation： GE Jianyun,CAI Haibo,DU Zheng,TAN Wensong.. EXPRESSIONS OF B-CELL-SPECIFIC MONOCLONAL LEUKEMIA VIRUS INSERT SITE 1 AND HUMAN TELOMERASE REVERSE TRANSCRIPTASE GENES IN CD34+ CELLS DURING EX VIVO EXPANSION PROCESS. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(9): 1112-1116. doi: Copy

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2.	Martinez-Agosto JA, Mikkola HK, Hartenstein V, et al. The hematopoietic stem cell and its niche: a comparative view. Genes Dev, 2007, 21(23): 3044-3060.
3.	Jaroscak J, Goltry K, Smith A, et al. Augmentation of umbilical cord blood (UCB) transplantation with ex vivo-expanded UCB cells: results of a phase 1 trial using the AastromReplicell System. Blood, 2003, 101(12): 5061-5067.
4.	Giarratana MC, Rouard H, Dumont A, et al. Proof of principle for transfusion of in vitro-generated red blood cells. Blood, 2011, 118(19): 5071-5079.
5.	Ann D, Colleen D, Bernstein ID. Ex vivo expansion of human hematopoietic stem and progenitor cells. Blood, 2011, 117(23): 6083-6090.
6.	Hofmeister CC, Zhang J, Knight KL, et al. Ex vivo expansion of umbilical cord blood stem cells for transplantation: growing knowledge from the hematopoietic niche. Bone Marrow Transplant, 2007, 39(1): 11-23.
7.	Law P, Traylor L, Recktenwald DJ. Cell analysis for hematopoietic stem/progenitor cells transplantation. Cytometry, 1999, 38(2): 47-52.
8.	Raaphorst FM. Self-renewal of hematopoietic and leukemic stem cells: a central role for the Polycomb-group gene Bmi-1. Trends Immunol, 2003, 24(10): 522-524.
9.	Oguro H, Yuan J, Ichikawa H, et al. Poised lineage specification in multipotential hematopoietic stem and progenitor cells by the polycomb protein Bmi1. Cell Stem Cell, 2010, 6(3): 279-286.
10.	Wu KJ, Grandori C, Amacker M, et al. Direct activation of TERT transcription by c-MYC. Nat Genet, 1999, 21(2): 220-224.
11.	Engelhardt M, Kumar R, Albanell J, et al. Telomerase regulation, cell cycle, and telomere stability in primitive hematopoietic cells. Blood, 1997, 90(1): 182-193.
12.	Kawano Y, Kobune M, Yamaguchi M, et al. Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells. Blood, 2003, 101(2): 532-540.
13.	Mohty M, Yong AS, Szydlo RM, et al. The polycomb group BMI1 gene is a molecular marker for predicting prognosis of chronic myeloid leukemia. Blood, 2007, 110(1): 380-383.
14.	Fan Y, Liu Z, Fang X, et al. Differential expression of full-length telomerase reverse transcriptase mRNA and telomerase activity between normal and malignant renal tissues. Clin Cancer Res, 2005, 11(12): 4331-4337.
15.	Yao CL, Chu LM, Hsieh TB, et al. A systematic strategy to optimize ex vivo expansion medium for human hematopoietic stem cells derived from umbilical Cord blood mononuclear cells. Exp Hematol, 2004, 32(8): 720-727.
16.	Gluckman E. Current status of umbilical cord blood hematopoietic stem cell transplantation. Exp Hematol, 2000, 28(11): 1197-1205.
17.	Jacobs JJ, Scheijen B, Voncken JW, et al. Bmi-1 collaborates with c-Myc in tumorigenesis by inhibiting c-Myc-induced apoptosis via INK4a/ARF. Genes Dev, 1999, 13(20): 2678-2690.
18.	Iwama A, Oguro H, Negishi M, et al. Enhanced self-renewal of hematopoietic stem cells mediated by the polycomb gene product Bmi-1. Immunity, 2004, 21(6): 843-851.
19.	Dick JE. Stem cell concepts renew cancer research. Blood, 2008, 112(13): 4793-4807.

1. 1 Rubinstein P, Carrier C, Scaradavou A, et al. Outcomes among 562 recipients of placental-blood transplants from unrelated donors. N Engl J Med, 1998, 339(22): 1565-1577. 2 Forraz N, McGuckin CP. The umbilical cord: a rich and ethical stem cell source to advance regenerative medicine. Cell Prolif, 2011, 44 Suppl 1: 60-69.
2. Martinez-Agosto JA, Mikkola HK, Hartenstein V, et al. The hematopoietic stem cell and its niche: a comparative view. Genes Dev, 2007, 21(23): 3044-3060.
3. Jaroscak J, Goltry K, Smith A, et al. Augmentation of umbilical cord blood (UCB) transplantation with ex vivo-expanded UCB cells: results of a phase 1 trial using the AastromReplicell System. Blood, 2003, 101(12): 5061-5067.
4. Giarratana MC, Rouard H, Dumont A, et al. Proof of principle for transfusion of in vitro-generated red blood cells. Blood, 2011, 118(19): 5071-5079.
5. Ann D, Colleen D, Bernstein ID. Ex vivo expansion of human hematopoietic stem and progenitor cells. Blood, 2011, 117(23): 6083-6090.
6. Hofmeister CC, Zhang J, Knight KL, et al. Ex vivo expansion of umbilical cord blood stem cells for transplantation: growing knowledge from the hematopoietic niche. Bone Marrow Transplant, 2007, 39(1): 11-23.
7. Law P, Traylor L, Recktenwald DJ. Cell analysis for hematopoietic stem/progenitor cells transplantation. Cytometry, 1999, 38(2): 47-52.
8. Raaphorst FM. Self-renewal of hematopoietic and leukemic stem cells: a central role for the Polycomb-group gene Bmi-1. Trends Immunol, 2003, 24(10): 522-524.
9. Oguro H, Yuan J, Ichikawa H, et al. Poised lineage specification in multipotential hematopoietic stem and progenitor cells by the polycomb protein Bmi1. Cell Stem Cell, 2010, 6(3): 279-286.
10. Wu KJ, Grandori C, Amacker M, et al. Direct activation of TERT transcription by c-MYC. Nat Genet, 1999, 21(2): 220-224.
11. Engelhardt M, Kumar R, Albanell J, et al. Telomerase regulation, cell cycle, and telomere stability in primitive hematopoietic cells. Blood, 1997, 90(1): 182-193.
12. Kawano Y, Kobune M, Yamaguchi M, et al. Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT)-transfected human stromal cells. Blood, 2003, 101(2): 532-540.
13. Mohty M, Yong AS, Szydlo RM, et al. The polycomb group BMI1 gene is a molecular marker for predicting prognosis of chronic myeloid leukemia. Blood, 2007, 110(1): 380-383.
14. Fan Y, Liu Z, Fang X, et al. Differential expression of full-length telomerase reverse transcriptase mRNA and telomerase activity between normal and malignant renal tissues. Clin Cancer Res, 2005, 11(12): 4331-4337.
15. Yao CL, Chu LM, Hsieh TB, et al. A systematic strategy to optimize ex vivo expansion medium for human hematopoietic stem cells derived from umbilical Cord blood mononuclear cells. Exp Hematol, 2004, 32(8): 720-727.
16. Gluckman E. Current status of umbilical cord blood hematopoietic stem cell transplantation. Exp Hematol, 2000, 28(11): 1197-1205.
17. Jacobs JJ, Scheijen B, Voncken JW, et al. Bmi-1 collaborates with c-Myc in tumorigenesis by inhibiting c-Myc-induced apoptosis via INK4a/ARF. Genes Dev, 1999, 13(20): 2678-2690.
18. Iwama A, Oguro H, Negishi M, et al. Enhanced self-renewal of hematopoietic stem cells mediated by the polycomb gene product Bmi-1. Immunity, 2004, 21(6): 843-851.
19. Dick JE. Stem cell concepts renew cancer research. Blood, 2008, 112(13): 4793-4807.

Chinese Journal of Reparative and Reconstructive Surgery

EXPRESSIONS OF B-CELL-SPECIFIC MONOCLONAL LEUKEMIA VIRUS INSERT SITE 1 AND HUMAN TELOMERASE REVERSE TRANSCRIPTASE GENES IN CD34+ CELLS DURING EX VIVO EXPANSION PROCESS

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