[Long-term culture of plasma cells from patients with myeloma and establishment of a permanent cell line].

Sbornik lekarsky Pub Date : 2002-01-01
P Stöckbauer, I Spicka, P Otevrelová, J Straub, M Hradcová, P Klener
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Abstract

Unlabelled: Multiple myeloma plasma cells are actively dividing cells with the long surviving ability in the ex-vivo culture. In the effort for better understanding of the proliferative potential of malignant myeloma cells and establishment of permanent myeloma cell lines we performed long term cultures of human myeloma cells ex-vivo. During the last two years we cultured 41 bone marrow samples from 39 patients with multiple myeloma. Cells were cultured in the RPMI 1640 culture medium with 15% fetal calf serum at 37 degrees C in 5% CO2 and approximately one third of the culture medium was changed regularly twice a week. Most of the marrows cultures died by apoptosis within 30 days. Four bone marrow samples were cultured for more than 11 months, however, no culture can be qualified as an established cell line. In three cases permanent B-lymphoblastoid cell lines were established (UHKT-55, UHKT-56 a UHKT-57) but secondary immortalization by Epstein-Barr virus was suggested.

Conclusions: Presented results suggest that myeloma plasma cells can survive and are able to proliferate in the ex-vivo culture for several months up to one year independently on the addition of any external growth factor without spontaneous apoptosis or necrosis. The probability of the establishment of a permanent cell line of plasma cell origin is, however, low. Presence of accessory bone marrow cells was the most important factor for the long-term survival of myeloma cells.

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[骨髓瘤患者浆细胞的长期培养和永久细胞系的建立]。
未标记:多发性骨髓瘤浆细胞是活跃分裂的细胞,在离体培养中具有较长的存活能力。为了更好地了解恶性骨髓瘤细胞的增殖潜力和建立永久性骨髓瘤细胞系,我们对人类骨髓瘤细胞进行了离体长期培养。在过去的两年中,我们培养了39例多发性骨髓瘤患者的41个骨髓样本。细胞在含有15%胎牛血清的RPMI 1640培养基中培养,37℃,5% CO2,大约三分之一的培养基每周定期更换两次。大多数骨髓培养物在30天内凋亡死亡。4个骨髓样本的培养时间超过11个月,但没有一个培养物可以作为已建立的细胞系。在三个病例中,建立了永久性b淋巴母细胞样细胞系(UHKT-55、UHKT-56和UHKT-57),但建议用eb病毒进行二次永生化。结论:目前的研究结果表明,骨髓瘤浆细胞可以独立地在体外培养中存活数月至一年,并且能够在添加任何外部生长因子的情况下增殖,而不会自发凋亡或坏死。然而,建立浆细胞来源的永久细胞系的可能性很低。辅助骨髓细胞的存在是骨髓瘤细胞长期存活的最重要因素。
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