Title: Ex-vivo expansion of nonhuman primate CD34+ cells by stem cell factor Sall4B.
Authors: Shen, Bin; Zhang, Yu; Dai, Wei; Ma, Yupo; Jiang, Yongping
Published In Stem Cell Res Ther, (2016 10 20)
Abstract: Hematopoietic CD34+ stem cells are widely used in the clinical therapy of complicated blood diseases. Stem cell factor Sall4B is a zinc finger transcription factor that plays a vital role in hematopoietic stem cell expansion. The purpose of our current study is to further evaluate how Sall4B might affect the expansion of CD34+ cells derived from nonhuman primates.Sall4B was overexpressed in nonhuman primate bone marrow-derived CD34+ cells via a lentiviral transduction system. The granulocyte-erythrocyte-macrophage-megakaryocyte colony-forming unit (CFU) assay evaluated the differentiation potential of primate CD34+ cells that were expanded with Sall4B. Furthermore, an in-vivo murine system was employed to evaluate the hematopoietic potential of primate Sall4B-expanded CD34+ cells.Overexpression of Sall4B promoted ex-vivo nonhuman primate CD34+ cell expansion by 9.21 ± 1.94-fold on day 9, whereas lentiviral transduction without Sall4B expanded cells by only 2.95 ± 0.77-fold. Sall4B maintained a significant percentage of CD34+ cells as well. The CFU assay showed that the Sall4B-expanded CD34+ cells still possessed multilineage differentiation potential. A study using nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice in vivo revealed that Sall4B led to an increase in the number of repopulating cells and the 9-day-old Sall4B-transduced CD34+ cells still possess self-renewal and multilineage differentiation capacity in vivo, which are similar stemness characteristics to those in freshly isolated primate bone marrow-derived CD34+ cells.We investigated the expansion of nonhuman primate bone marrow-derived CD34+ cells using the Sall4B lentiviral overexpression approach; our findings provide a new perspective on mechanisms of rapid stem cell proliferation. The utilization of Sall4B to expand CD34+ cells on a large scale through use of suitable model systems would prove helpful towards preclinical trials of autologous transplantation.
PubMed ID: 27765075
MeSH Terms: Animals; Antigens, CD34/metabolism*; Cell Differentiation/physiology; Cell Line; Cell Proliferation/physiology; Colony-Forming Units Assay/methods; Female; HEK293 Cells; Hematopoietic Stem Cell Transplantation/methods; Hematopoietic Stem Cells/metabolism; Humans; Macaca fascicularis; Male; Mice; Mice, Inbred NOD; Mice, SCID; Primates; Stem Cell Factor/metabolism*; Transcription Factors/metabolism*; Transplantation, Autologous/methods