Bachier C R, Gokmen E, Teale J, Lanzkron S, Childs C, Franklin W, Shpall E, Douville J, Weber S, Muller T, Armstrong D, LeMaistre C F
South Texas Cancer Institute, San Antonio 77229, USA.
Exp Hematol. 1999 Apr;27(4):615-23. doi: 10.1016/s0301-472x(98)00085-x.
The use of hematopoietic growth factors, stromal monolayers, and frequent medium exchange allows the expansion of hematopoietic progenitors ex-vivo. We evaluated the use of ex-vivo expanded progenitor cells for hematopoietic reconstitution following high dose chemotherapy (HDC) in breast cancer patients. Patients with high-risk Stage II or metastatic breast carcinoma underwent bone marrow aspirations using general anesthesia. A total of 675-1125 x 10(6) mononuclear cells (MNC) were seeded for ex-vivo expansion for 12 days in controlled perfusion bioreactors (Aastrom Biosciences, Inc.). The bone marrow cultures, which included the stromal cells collected with the aspirate, were supplemented with erythropoietin, granulocyte-macrophage-colony stimulating factor (GM-CSF)/IL-3 fusion protein (PIXY 321), and flt3 ligand. Stem cell transplant was performed with expanded cells after HDC. A median bone marrow volume of 52.9 mL (range 42-187 mL) was needed to inoculate the bioreactors. Median fold expansion of nucleated cells (NC) and colony forming unit granulocyte-macrophage (CFU-GM) was 4.9 and 9.5, respectively. The median fold expansion of CD34+lin- and long-term culture-initiating culture (LTC-IC) was 0.42 and 0.32, respectively. Five patients were transplanted with ex-vivo expanded NC. Median days to an absolute neutrophil count > 500/microL was 18 (range 15-22). Median days to a platelet count > 20,000/microl was 23 (range 19-39). All patients had sustained engraftment of both neutrophils and platelets. Immune reconstitution was similar to that seen after HDC and conventional stem cell transplantation. We conclude that ex-vivo expansion of progenitor cells from perfusion cultures of small volume bone marrow aspirates, allows hematopoietic reconstitution after HDC.
造血生长因子、基质单层培养以及频繁更换培养基的方法可实现造血祖细胞的体外扩增。我们评估了在乳腺癌患者接受大剂量化疗（HDC）后，使用体外扩增的祖细胞进行造血重建的效果。高危II期或转移性乳腺癌患者在全身麻醉下进行骨髓穿刺。将总共675 - 1125×10⁶个单核细胞（MNC）接种到可控灌注生物反应器（Aastrom Biosciences公司）中进行12天的体外扩增。骨髓培养物（包括穿刺采集的基质细胞）添加了促红细胞生成素、粒细胞 - 巨噬细胞集落刺激因子（GM - CSF）/IL - 3融合蛋白（PIXY 321）以及flt3配体。在HDC后用扩增后的细胞进行干细胞移植。接种生物反应器所需的骨髓体积中位数为52.9 mL（范围42 - 187 mL）。有核细胞（NC）和集落形成单位粒细胞 - 巨噬细胞（CFU - GM）的扩增倍数中位数分别为4.9和9.5。CD34⁺lin⁻和长期培养起始细胞（LTC - IC）的扩增倍数中位数分别为0.42和0.32。5例患者接受了体外扩增的NC移植。绝对中性粒细胞计数>500/μL的中位天数为18天（范围15 - 22天）。血小板计数>20,000/μL的中位天数为23天（范围19 - 39天）。所有患者的中性粒细胞和血小板均实现了持续植入。免疫重建情况与HDC和传统干细胞移植后的情况相似。我们得出结论，从小体积骨髓穿刺物的灌注培养物中体外扩增祖细胞，能够在HDC后实现造血重建。
