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美国科学家的最新研究表明Rho激酶抑制剂(Y-27632)与纤维母细胞饲养细胞合用能够诱导许多组织的正常上皮细胞以及肿瘤上皮细胞在体外无限增值,而不需要转导外源性病毒或细胞基因。例如原始前列腺和乳腺细胞重编程为基底细胞样干细胞样表型并在基底膜中形成有序的前列腺球和乳腺球。相较于筛选稀少的干细胞样细胞,在所述的生长条件下能够在活检组织获取后的5 d～6 d内产生2×10~6个细胞,能够从冷冻组织少于4个活细胞培养起来。持续的细胞增殖依赖于纤维母细胞饲养细胞和Y-27632两者,进行有条件重组的细胞(CRCs)保持了正常的核型和非致瘤性。该技术也能够有效建立人和啮齿类动物肿瘤细胞的培养系。例如,从人前列腺癌建立的CRCs表现出第13号染色体的不稳定性,在基底膜中的异常增殖以及在有严重合并免疫缺陷的老鼠中形成肿瘤。这种从少量活检样本和冷冻组织迅速产生许多肿瘤细胞的能力为基于细胞的诊断和治疗(包括测试化学敏感性)提供了很大机会并大大提高了生物银行的价值。另外,CRC方法能够对取自体内的上皮细胞进行遗传操作以及随后的自体体内评估。
Recent studies by U.S. scientists show that combination of Rho kinase inhibitor (Y-27632) with fibroblast feeder cells induces unlimited proliferation of normal epithelial cells and tumor epithelial cells in many tissues in vitro without the need to transduce exogenous viruses or cells gene. For example, prostatic and mammary gland cells are reprogrammed to a basal-like stem cell-like phenotype and form ordered prostatic and mammary glands in the basement membrane. Compared to the less screened stem cell-like cells, 2 × 10 ~ 6 cells could be produced within 5 d ~ 6 d after the biopsy tissue was obtained under the above-mentioned growth conditions, and less than 4 viable cells could be cultured from the frozen tissue stand up. Sustained cell proliferation depends on both fibroblast feeder cells and Y-27632, and cells that undergo conditional recombination (CRCs) retain normal karyotype and non-tumorigenicity. This technique is also effective in establishing culture lines for human and rodent tumor cells. For example, CRCs established from human prostate cancer exhibit instability of chromosome 13, aberrant proliferation in the basement membrane, and formation of tumors in mice with severe combined immunodeficiency. This ability to rapidly generate many tumor cells from a small number of biopsied samples and frozen tissue offers great opportunities for cell-based diagnosis and treatment, including testing for chemosensitivity, and greatly increases the value of bio-banking. In addition, the CRC method enables genetic manipulation of epithelial cells taken from the body and subsequent in vivo evaluation.