FUNCTIONALLY DISTINCT HUMAN MARROW STROMAL CELL-LINES IMMORTALIZED BY TRANSDUCTION WITH THE HUMAN PAPILLOMA-VIRUS E6/E7 GENES
FUNCTIONALLY DISTINCT HUMAN MARROW STROMAL CELL-LINES IMMORTALIZED BY TRANSDUCTION WITH THE HUMAN PAPILLOMA-VIRUS E6/E7 GENES
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DOI：
10.1182/blood.v85.4.997.bloodjournal854997
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发表时间：
1995-02-15
期刊：
影响因子：
20.3
通讯作者：
TOROKSTORB, B
中科院分区：
文献类型：
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作者：
ROECKLEIN, BA;TOROKSTORB, B
用含有人乳头瘤病毒E6/E7基因的复制缺陷型重组逆转录病毒（LXSN-16 E6 E7）使人骨髓基质细胞永生化。E6/E7基因产物分别干扰肿瘤抑制蛋白p53和Rb的功能，从而阻止细胞周期停滞而不引起显著的转化。27个永生化的克隆命名为HS-1 HS-27分离，其中四个的特点在本报告中。两种细胞系HS-5和HS-21似乎是成纤维细胞样的，并分泌显著水平的粒细胞集落刺激因子（G-CSF）、粒细胞-巨噬细胞-CSF（GM-CSF）、巨噬细胞-CSF（M-CSF）、Kit配体（KL）、巨噬细胞抑制蛋白-1 α、白细胞介素-6（IL-6）、IL-8和IL-11。然而，只有HS-5支持造血祖细胞增殖时，共培养在血清剥夺培养基没有外源性因素。与含有10 ng/mL IL-1、IL-3、IL-6、G-CSF、GM-CSF和KL以及3 U促红细胞生成素（Epo）的重组因子混合物相比，来自HS-5的条件培养基（CM）促进髓系集落生长的程度显著更大。另外两个克隆，HS-23和HS-27，类似于“毯子”细胞，具有上皮样形态，与HS-5和HS-21相比，更大，更宽，更平坦。这些细胞系分泌低水平的生长因子，并且不支持共培养物中分离的祖细胞的增殖。来自HS-23和HS-27的CM也不能支持骨髓集落的生长。HS-23和HS-27都表达相对高水平的VCAM-1，但HS-27是唯一支持分离的CD 34（+）38（lo）细胞形成“鹅卵石"区域的细胞系。我们假设HS-5、HS-21、HS-23和HS-27代表骨髓微环境的功能不同的组分。(C)1995年，美国血液学会。
A replication-defective recombinant retrovirus containing the human papilloma virus E6/E7 genes (LXSN-16 E6E7) was used to immortalize stromal cells from human marrow. The E6/E7 gene products interfere with the function of tumor-suppressor proteins p53 and Rb, respectively, thereby preventing cell cycle arrest without causing significant transformation. Twenty-seven immortalized clones designated HS-1 to HS-27 were isolated, four of which are characterized in this report. Two cell lines, HS-5 and HS-21, appear to be fibroblastoid and secrete significant levels of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-CSF (GM-CSF), macrophage-CSF (M-CSF), Kit ligand (KL), macrophage-inhibitory protein-1 alpha, interleukin-6 (IL-6), IL-8, and IL-11. However, only HS-5 supports proliferation of hematopoietic progenitor cells when cocultured in serum-deprived media with no exogenous factors. Conditioned media (CM) from HS-5 promotes growth of myeloid colonies to significantly greater extent than a cocktail of recombinant factors containing 10 ng/mL of IL-1, IL-3, IL-6, G-CSF, GM-CSF, and KL and 3 U of erythropoietin (Epo). Two additional clones, HS-23 and HS-27, resemble ''blanket'' cells, with an epithelioid morphology, and are much larger, broader, and flatter when compared with HS-5 and HS-21. These lines secrete low levels of growth factors and do not support proliferation of isolated progenitor cells in cocultures. CM from HS-23 and HS-27 also fail to support growth of myeloid colonies. Both HS-23 and HS-27 express relatively high levels of VCAM-1, yet HS-27 is the only line that supports the formation of ''cobblestone'' areas by isolated CD34(+)38(lo) cells. We hypothesize that HS-5, HS-21, HS-23, and HS-27 represent functionally distinct components of the marrow microenvironment. (C) 1995 by The American Society of Hematology.