Potdar Pd, Subedi Rp
Head, department of molecular medicine and biology, Jaslok Hospital and Research centre , Mumbai, India.
Research Student, Jaslok Hospital and Research centre , Mumbai, India.
J Stem Cells Regen Med. 2011 Apr 1;7(1):29-40. doi: 10.46582/jsrm.0701004. eCollection 2011.
Acute Lymphocytic Leukemia (ALL) is a clonal myeloid disorder affecting all age groups, characterized by accumulation of immature blast cells in bone marrow and in peripheral blood. Autologous Bone Marrow Transplantation is a present treatment for cure of ALL patients, which is very expensive, invasive process and may have possibility of transplantation of malignant stem cells to patients. In the present study, we hypothesized to isolate large number of normal Mesenchymal & Hematopoietic stem cells from peripheral blood of ALL patients, which will be further characterized for their normal phenotypes by using specific molecular stem cell markers. This is the first study, which defines the existing phenotypes of isolated MSCs and HSCs from peripheral blood of ALL patients. We have established three cell lines in which two were Mesenchymal stem cells designated as MSCALL and MSCnsALL and one was suspension cell line designated as HSCALL. The HSCALL cell line was developed from the lymphocyte like cells secreted by MSCALL cells. Our study also showed that MSCALL from peripheral blood of ALL patient secreted hematopoietic stem cells in vitro culture. We have characterized all three-cell lines by 14 specific stem cell molecular markers. It was found that both MSC cell lines expressed CD105, CD13, and CD73 with mixed expression of CD34 and CD45 at early passage whereas, HSCALL cell line expressed prominent feature of hematopoietic stem cells such as CD34 and CD45 with mild expression of CD105 and CD13. All three-cell lines expressed LIF, OCT4, NANOG, SOX2, IL6, and DAPK. These cells mildly expressed COX2 and did not express BCR-ABL. Overall it was shown that isolated MSCs and HSCs can be use as a model system to study the mechanism of leukemia at stem cell level and their use in stem cell regeneration therapy for Acute Lymphocytic Leukemia.
急性淋巴细胞白血病（ALL）是一种影响所有年龄组的克隆性髓系疾病，其特征是未成熟的原始细胞在骨髓和外周血中积聚。自体骨髓移植是目前治疗ALL患者以实现治愈的方法，这是一个非常昂贵且具有侵入性的过程，并且可能存在将恶性干细胞移植给患者的可能性。在本研究中，我们假设从ALL患者的外周血中分离出大量正常的间充质和造血干细胞，并使用特定的分子干细胞标志物对其正常表型进行进一步表征。这是第一项定义从ALL患者外周血中分离出的间充质干细胞（MSCs）和造血干细胞（HSCs）现有表型的研究。我们建立了三种细胞系，其中两种是间充质干细胞，分别命名为MSCALL和MSCnsALL，一种是悬浮细胞系，命名为HSCALL。HSCALL细胞系是由MSCALL细胞分泌的淋巴细胞样细胞发育而来的。我们的研究还表明，ALL患者外周血中的MSCALL在体外培养中分泌造血干细胞。我们通过14种特定的干细胞分子标志物对这三种细胞系进行了表征。结果发现，两种MSC细胞系在早期传代时均表达CD105、CD13和CD73，同时混合表达CD34和CD45，而HSCALL细胞系表达造血干细胞的显著特征，如CD34和CD45，同时轻度表达CD105和CD13。所有三种细胞系均表达白血病抑制因子（LIF）、八聚体结合转录因子4（OCT4）、 Nanog同源物（NANOG）、性别决定区Y框蛋白2（SOX2）、白细胞介素6（IL6）和死亡相关蛋白激酶（DAPK）。这些细胞轻度表达环氧化酶2（COX2），不表达断裂簇区域-阿贝尔逊（BCR-ABL）。总体而言，研究表明，分离出的MSCs和HSCs可作为一种模型系统，用于在干细胞水平研究白血病的发病机制及其在急性淋巴细胞白血病干细胞再生治疗中的应用。
