Ex-vivo expansion of nonhuman primate CD34+ cells by stem cell factor Sall4B.

Abstract

BACKGROUND:

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.

METHODS:

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.

RESULTS:

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.

CONCLUSIONS:

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.

• Authors: Bin Shen, Yu Zhang, Wei Dai, Yupo Ma, Yongping Jiang
• Journal: Stem cell research & therapy (Stem Cell Res Ther) Vol. 7 Issue 1 Pg. 152 (10 20 2016) ISSN: 1757-6512 [Electronic] England
• PMID: 27765075 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, CD34
• Stem Cell Factor
• Transcription Factors

Topics

• 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)