Human adipose tissue‐derived stromal cells can differentiate into megakaryocytes and platelets by secreting endogenous thrombopoietin

Abstract

EssentialsManufacturing platelets from a donor-independent source is highlighted in transfusion medicine. We examined the differentiation of adipose tissuederived stromal cells (ASCs) into platelets. Endogenous thrombopoietin (TPO) induced ASCs differentiation into megakaryocytes and platelets. TPO secretion from ASCs was due to an interaction of transferrin with its receptor CD71.Summary. Background: Ex vivo production of megakaryocytes (MKs) and platelets from a donor-independent source is currently of intense interest in transfusion medicine. Adipose tissue-derived stromal cells (ASCs) constitute an attractive candidate cell source, because inducing these cells into MK lineages requires no gene transfer and only endogenous transcription factors containing p45NF-E2/Maf, an MK-inducing factor. Objectives: To examine whether ASCs differentiate into MK lineages by using endogenous thrombopoietin (TPO), a primary cytokine that drives MK lineages. Methods: TPO levels were measured by quantitative real-time PCR and ELISA. To investigate the effects of endogenous TPO on MK and platelet production, surface marker expression and functions for platelets were analyzed in ASC-derived cells cultured in the presence or absence of recombinant TPO. Based on a screening test, the role of transferrin receptor CD71 in TPO production and MK differentiation was examined with anti-CD71 antibody, small interfering RNA (siRNA) against CD71 (siRNA-CD71), and CD71-positive/negative cells. Results: ASCs secreted TPO during MK differentiation, and the endogenous TPO facilitated MK and platelet production from ASCs. TPO secretion from ASCs occurred in a transferrin-dependent manner. ASCs treated with anti-CD71 antibody or transfected with siRNA-CD71 produced markedly less TPO. The TPO levels and MK yield were significantly higher when CD71-positive ASCs were used than when CD71-negative ASCs were used. Conclusions: CD71 might be an appropriate marker for MK progenitor cells among human ASCs, because of the higher capacity of CD71-positive cells to produce TPO and their ability to differentiate into MKs. These findings could help to establish an efficient method for platelet production.

Platelets in liver disease, cancer and regeneration

“…Furthermore, Ono-Uruga et al[86] have discovered that pre-adipocytes endogenously possess the megakaryocyte inducer p45NF-E2 and can induce differentiation of megakaryocytes into platelets while increasing their own expression of p45NF-E2; they are striving to stably and safely produce platelets for transfusion from pre-adipocytes for medical applications. However, the number of platelets produced by this method is currently too low for practical use.…”

Adipose-derived stromal cells in regulation of hematopoiesis

“…However, Zhu et al suggested that ASCs inhibited the proliferation of erythroleukemia K562 cells in vitro [29]. It has also been proved that ASCs can promote development of megakaryocytes and platelets [30]. The recent studies have not drawn a consistent conclusion on hematopoietic regulation biases of ASCs.…”

Current status of blood ‘pharming’: megakaryoctye transfusions as a source of platelets

“…Finally, alternative adult tissues have been used to generate megakaryocytes and platelets. While endothelial cells may give rise to megakaryocytes under appropriate stimulation [34], the most successful approach begins with liposuction-derived adipocytes that can be grown in an adipocyte-induction medium followed by differentiation in a megakaryocyte-induction media [35]. The advantages of the latter approach are that the necessary beginning cells are readily available and induction to produce megakaryocytes requires no genetic manipulation.…”