Papa Luena, Djedaini Mansour, Hoffman Ronald
Division of Hematology/Oncology, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai;
Division of Hematology/Oncology, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai.
J Vis Exp. 2019 Apr 11(146). doi: 10.3791/59532.
Umbilical cord blood (UCB) units provide an alternative source of human hematopoietic stem cells (HSCs) for patients who require allogeneic bone marrow transplantation. While UCB has several unique advantages, the limited numbers of HSCs within each UCB unit limits their use in regenerative medicine and HSC transplantation in adults. Efficient expansion of functional human HSCs can be achieved by ex vivo culturing of CD34 cells isolated from UCBs and treated with a deacetylase inhibitor, valproic acid (VPA). The protocol detailed here describes the culture conditions and methodology to rapidly isolate CD34 cells and expand to a high degree a pool of primitive HSCs. The expanded HSCs are capable of establishing both short-term and long-term engraftment and are able to give rise to all types of differentiated hematopoietic cells. This method also holds potential for clinical application in autologous HSC gene therapy and provides an attractive approach to overcome the loss of functional HSCs associated with gene editing.
脐带血（UCB）单位为需要异基因骨髓移植的患者提供了人类造血干细胞（HSC）的替代来源。虽然脐带血有几个独特的优点，但每个脐带血单位内造血干细胞数量有限，限制了它们在再生医学和成人造血干细胞移植中的应用。通过体外培养从脐带血中分离出的CD34细胞并用去乙酰化酶抑制剂丙戊酸（VPA）处理，可以实现功能性人类造血干细胞的有效扩增。此处详细描述的方案介绍了快速分离CD34细胞并高度扩增原始造血干细胞库的培养条件和方法。扩增后的造血干细胞能够实现短期和长期植入，并能够产生所有类型的分化造血细胞。该方法在自体造血干细胞基因治疗中也具有临床应用潜力，并为克服与基因编辑相关的功能性造血干细胞损失提供了一种有吸引力的方法。
