Gudleviciene Zivile, Kundrotas Gabrielis, Liudkeviciene Regina, Rascon Jelena, Jurga Marcin
National Cancer Institute, Biobank, Santariskiu 1, 08660 Vilnius, Lithuania, tel. +37052190909, fax. +37052720164.
National Cancer Institute, Biobank Santariskiu 1, 08660 Vilnius, Lithuania Vilnius University, Faculty of Natural Sciences M.K. Ciurlionio 21/27, 03101 Vilnius.
Open Med (Wars). 2014 Oct 8;10(1):44-49. doi: 10.1515/med-2015-0008. eCollection 2015.
Mesenchymal stem cells (MSCs) are currently exploited in numerous clinical trials to investigate their potential in immune regulation, hematopoesis or tissue regeneration. The most common source of MSCs for clinical use is human bone marrow. To generate sufficient numbers of cells relevant to clinical use in most cases the high volumes (20-50 ml) of bone marrow aspirates are taken.
In this pilot study, 8 healthy bone marrow donors were included. Two different MSC extraction methods were evaluated: MSCs extraction from 60 ml of bone marrow using density gradient and MSCs extraction from 6 ml using red blood cell (RBC) lysis.
Our results showed that after RBC lysis the efficient amount of human MSCs can be isolated from 10 times less bone marrow volume (6 ml). Moreover, using small volume of bone marrow the adequate therapeutical dose of MSCs could be achieved during similar period of time (3-4 weeks). In conclusion, we have shown that MSCs isolation using RBC lysis is an effective and more advantageous method in comparison to standard MSCs isolation using density-gradient. Using RBC lysis from small volume of bone marrow the same amount of MSCs were obtained as usually using large volume and density-gradient.
间充质干细胞（MSCs）目前正在众多临床试验中被用于研究其在免疫调节、造血或组织再生方面的潜力。临床使用的MSCs最常见来源是人类骨髓。在大多数情况下，为了产生足够数量与临床使用相关的细胞，需要采集大量（20 - 50毫升）骨髓抽吸物。
在这项初步研究中，纳入了8名健康骨髓供体。评估了两种不同的MSCs提取方法：使用密度梯度从60毫升骨髓中提取MSCs以及使用红细胞（RBC）裂解从6毫升骨髓中提取MSCs。
我们的结果表明，经过RBC裂解后，可以从少10倍体积（6毫升）的骨髓中分离出有效数量的人类MSCs。此外，使用少量骨髓在相似时间段（3 - 4周）内可以获得足够的治疗剂量的MSCs。总之，我们已经表明，与使用密度梯度的标准MSCs分离方法相比，使用RBC裂解分离MSCs是一种有效且更具优势的方法。使用少量骨髓进行RBC裂解获得的MSCs数量与通常使用大量骨髓和密度梯度获得的数量相同。
