• 中國醫(yī)學(xué)科學(xué)院 北京協(xié)和醫(yī)院基本外科（北京 100730）;

目的探討胰腺癌細(xì)胞在二維平面培養(yǎng)和三維培養(yǎng)（Ⅰ型膠原和細(xì)胞外基質(zhì)膠）中的生長特性。方法將3株胰腺癌細(xì)胞株SW1990、PCT和ASPC1分別采用上述3種培養(yǎng)方式進(jìn)行培養(yǎng)，觀察細(xì)胞生長形態(tài)。采用CCK8法測定細(xì)胞生長曲線，采用乙醇固定碘化丙錠染色法檢測細(xì)胞周期分布。結(jié)果 細(xì)胞在二維平面培養(yǎng)系統(tǒng)中呈單層貼壁生長; 在Ⅰ型膠原和細(xì)胞外基質(zhì)膠中細(xì)胞形成多細(xì)胞球樣體（multicellular spheroid，MCS），其生長速度較二維平面培養(yǎng)中的細(xì)胞慢。在二維平面培養(yǎng)中生長的SW1990、PCT和ASPC1細(xì)胞的S期細(xì)胞的比例分別為(29.6±3.0)%、(33.6±2.1)%和(33.1±1.8)%，明顯高于在Ⅰ型膠原中培養(yǎng)4 d和8 d形成的MCS的S期細(xì)胞比例〔(18.2±5.1)%、(14.5±3.2)%和(24.7±2.6)%〕，P lt;0.05，而G2/M期的細(xì)胞比例差異沒有統(tǒng)計(jì)學(xué)意義（P gt;0.05）。在Ⅰ型膠原中培養(yǎng)4 d和8 d的SW1990和PCT細(xì)胞以及培養(yǎng)8 d的ASPC1細(xì)胞的G0/G1期細(xì)胞比例明顯高于其在二維平面培養(yǎng)中的細(xì)胞比例（P lt;0.05）。在Ⅰ型膠原中培養(yǎng)4 d的ASPC1細(xì)胞和SW1990細(xì)胞的S期細(xì)胞比例明顯高于其培養(yǎng)8 d的細(xì)胞比例（P lt;0.05）。結(jié)論不同的培養(yǎng)方式、培養(yǎng)介質(zhì)對胰腺癌細(xì)胞的生長具有較大的影響。MCS三維培養(yǎng)系統(tǒng)能更好地模擬體內(nèi)腫瘤細(xì)胞的生長狀況。

引用本文： 徐強(qiáng),胡亞,廖泉,趙玉沛. 胰腺癌細(xì)胞在二維和三維培養(yǎng)系統(tǒng)中的生長特性研究△. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(1): 33-38. doi: 復(fù)制

版權(quán)信息： ?四川大學(xué)華西醫(yī)院華西期刊社《中國普外基礎(chǔ)與臨床雜志》版權(quán)所有，未經(jīng)授權(quán)不得轉(zhuǎn)載、改編

1. Abbott A. Cell culture: biolog’s new dimension [J]. Nature, 2003, 424(6951): 870872.
2. Matsuura T. Bioreactors for 3dimensional highdensity culture of human cells [J]. Hum Cell, 2006, 19(1): 1116.
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4. Lee GY, Kenny PA, Lee EH, et al. Threedimensional culture models of normal and malignant breast epithelial cells [J]. Nat Methods, 2007, 4(4): 359365.
5. Lin RZ, Chang HY. Recent advances in threedimensional multicellular spheroid culture for biomedical research [J]. Biotechnol J, 2008, 3(910): 11721184.
6. 周濤, 胡亞, 廖泉, 等. 2種體外腫瘤細(xì)胞藥敏檢測方法的比較研究 [J]. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(11): 825828.
7. 周濤, 胡亞, 廖泉, 等. 胰腺癌多細(xì)胞團(tuán)簇的化療藥物敏感試驗(yàn) [J]. 中華胰腺病雜志, 2008, 8(6): 361364.
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10. MuellerKlieser W. Tumor biology and experimental therapeutics [J]. Crit Rev Oncol Hematol, 2000, 36(23): 123139.
11. Friedrich J, Ebner R, KunzSchughart LA. Experimental antitumor therapy in 3D: spheroids-old hat or new challenge? [J]. Int J Radiat Biol, 2007, 83(1112): 849871.
12. 周方正, 伍鋼. 細(xì)胞偽足與腫瘤轉(zhuǎn)移研究進(jìn)展 [J]. 臨床腫瘤學(xué)雜志, 2007, 12(1): 7779.
13. 張秀真, 吳澤志. 細(xì)胞偽足形成與微絲骨架研究進(jìn)展 [J]. 國外醫(yī)學(xué)臨床生物化學(xué)與檢驗(yàn)學(xué)分冊, 2005, 26(4): 213216.
14. 徐強(qiáng), 廖泉, 趙玉沛. 多細(xì)胞球樣體的制備及其在腫瘤治療研究中的應(yīng)用 [J]. 基礎(chǔ)醫(yī)學(xué)與臨床, 2010, 30(3): 320324.
15. Santini MT, Rainaldi G, Indovina PL. Multicellular tumour spheroids in radiation biology [J]. Int J Radiat Biol, 1999, 75(7): 787799.
16. Sminia P, Acker H, Eikesdal HP, et al. Oxygenation and response to irradiation of organotypic multicellular spheroids of human glioma [J]. Anticancer Res, 2003, 23(2B): 14611466.
17. Acker H, Carlsson J, MuellerKlieser W, et al. Comparative pO2 measurements in cell spheroids cultured with different techniques [J]. Br J Cancer, 1987, 56(3): 325327.
18. DomínguezMuoz JE著, 趙玉沛主譯. 臨床胰腺病學(xué) [M]. 天津: 天津科技翻譯出版公司, 2007: 292293.
19. 潘玉先, 鄭振華. 人大隱靜脈內(nèi)皮細(xì)胞種植人工血管的實(shí)驗(yàn)研究 [J]. 中國普外基礎(chǔ)與臨床雜志, 2000, 7(1): 1315.
20. 施森, 何延政, 劉勇, 等. 外周血來源內(nèi)皮祖細(xì)胞三維血管新生模型的建立及其特性分析 [J]. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(2): 119123.
21. Gen M, Castro Kreder N, Bartenvan Rijbroek A, et al. Enhancement of effects of irradiation by gemcitabine in a glioblastoma cell line and cell line spheroids [J]. J Cancer Res Clin Oncol, 2004, 130(1): 4551.
22. Muir CP, Adams MA, Graham CH. Nitric oxide attenuates resistance to doxorubicin in threedimensional aggregates of human breast carcinoma cells [J]. Breast Cancer Res Treat, 2006, 96(2): 169176.
23. Chua YJ, Zalcberg JR. Pancreatic cancer-is the wall crumbling? [J]. Ann Oncol, 2008, 19(7): 12241230.
24. Vulfovich M, RochaLima C. Novel advances in pancreatic cancer treatment [J]. Expert Rev Anticancer Ther, 2008, 8(6): 9931002.
25. Boeck S, Heinemann V. The role of secondline chemotherapy after gemcitabine failure in patients with advanced pancreatic cancer [J]. Future Oncol, 2008, 4(1): 4150.
26. Tempero M, Plunkett W, Ruiz Van Haperen V, et al. Randomized Phase Ⅱ comparison of doseintense gemcitabine: thirtyminute infusion and fixed dose rate infusion in patients with pancreatic adenocarcinoma [J]. J Clin Oncol, 2003, 21(18): 34023408.
27. Louvet C, Labianca R, Hammel P, et al. Gemcitabine in combination with oxaliplatin compared with gemcitabine alone in locally advanced or metastatic pancreatic cancer: results of a GERCOR and GISCAD phase Ⅲ trial [J]. J Clin Oncol, 2005, 23(15): 35093516.
  1. 1. Abbott A. Cell culture: biolog’s new dimension [J]. Nature, 2003, 424(6951): 870872.
  2. 2. Matsuura T. Bioreactors for 3dimensional highdensity culture of human cells [J]. Hum Cell, 2006, 19(1): 1116.
  3. 3. Del Duca D, Werbowetski T, Del Maestro RF. Spheroid preparation from hanging drops: characterization of a model of brain tumor invasion [J]. J Neurooncol, 2004, 67(3): 295303.
  4. 4. Lee GY, Kenny PA, Lee EH, et al. Threedimensional culture models of normal and malignant breast epithelial cells [J]. Nat Methods, 2007, 4(4): 359365.
  5. 5. Lin RZ, Chang HY. Recent advances in threedimensional multicellular spheroid culture for biomedical research [J]. Biotechnol J, 2008, 3(910): 11721184.
  6. 6. 周濤, 胡亞, 廖泉, 等. 2種體外腫瘤細(xì)胞藥敏檢測方法的比較研究 [J]. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(11): 825828.
  7. 7. 周濤, 胡亞, 廖泉, 等. 胰腺癌多細(xì)胞團(tuán)簇的化療藥物敏感試驗(yàn) [J]. 中華胰腺病雜志, 2008, 8(6): 361364.
  8. 8. Mellor HR, Ferguson DJ, Callaghan R. A model of quiescent tumor microregions for evaluating multicellular resistance to chemotherapeutic drugs [J]. Br J Cancer, 2005, 93(3): 302309.
  9. 9. KunzSchughart LA, Freyer JP, Hofstaedter F, et al. The use of 3D cultures for highthroughput screening: the multicellular spheroid model [J]. J Biomol Screen, 2004, 9(4): 273285.
  10. 10. MuellerKlieser W. Tumor biology and experimental therapeutics [J]. Crit Rev Oncol Hematol, 2000, 36(23): 123139.
  11. 11. Friedrich J, Ebner R, KunzSchughart LA. Experimental antitumor therapy in 3D: spheroids-old hat or new challenge? [J]. Int J Radiat Biol, 2007, 83(1112): 849871.
  12. 12. 周方正, 伍鋼. 細(xì)胞偽足與腫瘤轉(zhuǎn)移研究進(jìn)展 [J]. 臨床腫瘤學(xué)雜志, 2007, 12(1): 7779.
  13. 13. 張秀真, 吳澤志. 細(xì)胞偽足形成與微絲骨架研究進(jìn)展 [J]. 國外醫(yī)學(xué)臨床生物化學(xué)與檢驗(yàn)學(xué)分冊, 2005, 26(4): 213216.
  14. 14. 徐強(qiáng), 廖泉, 趙玉沛. 多細(xì)胞球樣體的制備及其在腫瘤治療研究中的應(yīng)用 [J]. 基礎(chǔ)醫(yī)學(xué)與臨床, 2010, 30(3): 320324.
  15. 15. Santini MT, Rainaldi G, Indovina PL. Multicellular tumour spheroids in radiation biology [J]. Int J Radiat Biol, 1999, 75(7): 787799.
  16. 16. Sminia P, Acker H, Eikesdal HP, et al. Oxygenation and response to irradiation of organotypic multicellular spheroids of human glioma [J]. Anticancer Res, 2003, 23(2B): 14611466.
  17. 17. Acker H, Carlsson J, MuellerKlieser W, et al. Comparative pO2 measurements in cell spheroids cultured with different techniques [J]. Br J Cancer, 1987, 56(3): 325327.
  18. 18. DomínguezMuoz JE著, 趙玉沛主譯. 臨床胰腺病學(xué) [M]. 天津: 天津科技翻譯出版公司, 2007: 292293.
  19. 19. 潘玉先, 鄭振華. 人大隱靜脈內(nèi)皮細(xì)胞種植人工血管的實(shí)驗(yàn)研究 [J]. 中國普外基礎(chǔ)與臨床雜志, 2000, 7(1): 1315.
  20. 20. 施森, 何延政, 劉勇, 等. 外周血來源內(nèi)皮祖細(xì)胞三維血管新生模型的建立及其特性分析 [J]. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(2): 119123.
  21. 21. Gen M, Castro Kreder N, Bartenvan Rijbroek A, et al. Enhancement of effects of irradiation by gemcitabine in a glioblastoma cell line and cell line spheroids [J]. J Cancer Res Clin Oncol, 2004, 130(1): 4551.
  22. 22. Muir CP, Adams MA, Graham CH. Nitric oxide attenuates resistance to doxorubicin in threedimensional aggregates of human breast carcinoma cells [J]. Breast Cancer Res Treat, 2006, 96(2): 169176.
  23. 23. Chua YJ, Zalcberg JR. Pancreatic cancer-is the wall crumbling? [J]. Ann Oncol, 2008, 19(7): 12241230.
  24. 24. Vulfovich M, RochaLima C. Novel advances in pancreatic cancer treatment [J]. Expert Rev Anticancer Ther, 2008, 8(6): 9931002.
  25. 25. Boeck S, Heinemann V. The role of secondline chemotherapy after gemcitabine failure in patients with advanced pancreatic cancer [J]. Future Oncol, 2008, 4(1): 4150.
  26. 26. Tempero M, Plunkett W, Ruiz Van Haperen V, et al. Randomized Phase Ⅱ comparison of doseintense gemcitabine: thirtyminute infusion and fixed dose rate infusion in patients with pancreatic adenocarcinoma [J]. J Clin Oncol, 2003, 21(18): 34023408.
  27. 27. Louvet C, Labianca R, Hammel P, et al. Gemcitabine in combination with oxaliplatin compared with gemcitabine alone in locally advanced or metastatic pancreatic cancer: results of a GERCOR and GISCAD phase Ⅲ trial [J]. J Clin Oncol, 2005, 23(15): 35093516.

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