In vivo generation of β-cell–like cells from CD34+ cells differentiated from human embryonic stem cells

Abstract

Objective-CD34 + cells, present within the bone marrow, have previously been shown to possess pancreatic endocrine potential. Based on this observation, we explored the capacity of CD34 + cells derived in culture from the differentiation of human embryonic stem cells (hESC), for their in-vivo pancreatic endocrine capacity.Methods-Sheep were transplanted with hESC-derived CD34 + cells, as well as non-sorted differentiated cultures. Transplantations were carried out with in-utero intraperitoneal injections prior to the development of the immune system in the fetus so that tolerance towards foreign antigens was acquired during gestation and persisted in the adult.Results-All cell populations that were tested demonstrated human cellular activity and long-term presence up to 5 years. However, the in-vivo beta-cell-like activity achieved from the transplantation of the sorted CD34 + cell population was not augmented by transplanting the entire cell population from which the CD34 + cells were isolated. Human DNA and insulin mRNA were detected in sheep pancreases. An average of 1.51 ng/mL human C-peptide was detected in serum from 8 animals transplanted with differentiated cell populations and assayed up to 55 months post-transplantation. Transplantation of as few as 23,500 cells resulted in long-term sustainable beta-cell like activity. Teratomas were absent in the transplanted animals.Conclusion-Our data suggest that hESC-derived CD34 + cells have a potential for long term invivo endocrine cellular activity which could prove useful in regenerative medicine. Since the same cell population has previously been shown to contain hematopoietic potential, it could be used for the induction of immunological tolerance and bone marrow chimerism prior to cellular therapy for diabetes. KeywordsHuman embryonic stem cells; Pancreas; Transplantation; Beta-cells; C-peptide Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. One potential therapy for diabetics is the restoration of beta-cell mass for the endogenous control of glycemia [1,2]. To this end, human embryonic stem cells (hESC) which have the capacity to form tissues of all three germ layers cells [3], are being evaluated for cellular transplantation therapy. hESC differentiation cultures have been developed for the derivation of progenitor cells that engraft and differentiate in-vivo into beta-cell-like cells [4,5]. Such research has had to occur in the absence of knowledge of a single or a group of cell-surface markers that may be used to unequivocally identify and isolate pancreatic stem cells [6]. Hence, focus has been placed ...

The CD34 surface antigen is restricted to glucagon-expressing cells in the early developing bovine pancreas

“…One could speculate that CD133/CD34 expression might begin before this point, but evidence supporting this is lacking and needs further investigation. The possibility of restoring lost beta cells by the reprogramming of alpha cells in vivo (Sangan and Tosh 2010;Thorel et al 2010), or by in vitro differentiation form CD34-positive cells derived from human embryonic stem cells (Goodrich et al 2010), is a tantalising thought. CD133/CD34-positive alpha cells may represent a defined source from which insulin-producing cells could be generated, and this is worthy of further study.…”

Influence of a dual-injection regimen, plerixafor and CXCR4 on in utero hematopoietic stem cell transplantation and engraftment with use of the sheep model

“…Tissue sections were mounted and baked onto slides. Target retrieval using citrate buffer was done as described previously (31). Immunohistochemistry (IHC) was carried out using rabbit anti-SDF1 antibody (clone RB32982) which reacted with both human and sheep tissue sections (Abgent, San Diego, CA), and/or mouse anti-human nuclei antibody (clone 235-1) (PhosphoSolutions, Aurora, CO) which only reacted with human cells.…”

Recent progress in histochemistry and cell biology

“…The present study demonstrated that CD34-and/or CD133-reactive cells contribute to the pancreatic alpha cell population during early fetal development in cattle. It has been previously shown that loss of beta cells can be replaced by alpha cells in vivo (Sangan and Tosh 2010;Thorel et al 2010), or by in vitro differentiation form CD34-positive cells derived from human embryonic stem cells (Goodrich et al 2010). Thus CD133/CD34-positive alpha cells may represent a defined source for insulinproducing cells.…”