• 作者單位：華東理工大學(xué)生物反應(yīng)器工程國(guó)家重點(diǎn)實(shí)驗(yàn)室（上海，200237）;

目的 研究體外擴(kuò)增過(guò)程中，CD34+細(xì)胞內(nèi)B細(xì)胞特異單克隆鼠白血病毒整合位點(diǎn)基因1（B-cell-specific monoclonal leukemia virus insert site 1，Bmi1）和人端粒酶逆轉(zhuǎn)錄酶（human telomerase reverse transcriptase，hTERT）基因的表達(dá)水平與其擴(kuò)增特性的相關(guān)性。 方法采用含F(xiàn)BS、干細(xì)胞生長(zhǎng)因子、Flt-3配體和促血小板生成素的IMDM培養(yǎng)基體外培養(yǎng)臍血CD34+細(xì)胞，在28 d培養(yǎng)過(guò)程中檢測(cè)CD34+細(xì)胞擴(kuò)增倍數(shù)、CD34+細(xì)胞比生長(zhǎng)速率以及集落形成率的變化趨勢(shì)，并采用熒光定量PCR檢測(cè)體外擴(kuò)增過(guò)程中CD34+細(xì)胞內(nèi)Bmi1和hTERT基因表達(dá)水平變化，分析以上基因表達(dá)水平與CD34+細(xì)胞擴(kuò)增特性之間的關(guān)系。 結(jié)果經(jīng)過(guò)28 d的體外培養(yǎng)，CD34+細(xì)胞共擴(kuò)增了（20.1 ± 3.5）倍，其占擴(kuò)增后總細(xì)胞的比例由培養(yǎng)前的95.5% ± 2.6%下降至2.1% ± 0.4%；CD34+細(xì)胞的比生長(zhǎng)速率和集落形成率均在培養(yǎng)7 d后出現(xiàn)明顯下降，而細(xì)胞內(nèi)Bmi1和hTERT mRNA的表達(dá)水平在培養(yǎng)7 d時(shí)達(dá)最高，此后逐漸下降至培養(yǎng)前水平。 結(jié)論Bmi1和hTERT基因表達(dá)與CD34+細(xì)胞體外增殖能力可能存在一定相關(guān)性。

引用本文： 葛劍云,蔡海波,杜錚,譚文松. 體外擴(kuò)增過(guò)程中CD34+細(xì)胞內(nèi)B細(xì)胞特異單克隆鼠白血病毒整合位點(diǎn)基因1和人端粒酶逆轉(zhuǎn)錄酶基因的表達(dá). 中國(guó)修復(fù)重建外科雜志, 2012, 26(9): 1112-1116. doi: 復(fù)制

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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. 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. 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. 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. 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. 5. Ann D, Colleen D, Bernstein ID. Ex vivo expansion of human hematopoietic stem and progenitor cells. Blood, 2011, 117(23): 6083-6090.
  6. 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. 7. Law P, Traylor L, Recktenwald DJ. Cell analysis for hematopoietic stem/progenitor cells transplantation. Cytometry, 1999, 38(2): 47-52.
  8. 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. 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. 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. 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. 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. 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. 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. 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. 16. Gluckman E. Current status of umbilical cord blood hematopoietic stem cell transplantation. Exp Hematol, 2000, 28(11): 1197-1205.
  17. 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. 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. 19. Dick JE. Stem cell concepts renew cancer research. Blood, 2008, 112(13): 4793-4807.