Title:

Ex vivo expansion and differentiation of haematopoietic stem cells to monocytes/macrophages

Author:

Stec, Małgorzata

Subject:

CD34+ cells ; macrophages ; monocytes ; expansion

Abstract:

The aim of this study was to design the methodology of expansion and differentiation of CD34+ cells to monocytes/macrophages. The present study shows that optimal medium for expansion of CD34+ cells was X-VIVO 10 supplemented with FBS, SCF, IL-3, FLT-3L, TPO and 3-4 days culture. For differentiation of CD34+ cells to monocytes/macrophages was IMDM supplemented with FBS, M-CSF, SCF, IL-3, FLT-3L and culture for 7-10 days. Using this protocol it was possible to obtain 1000 - fold increase in number of expanded cells, of which 40-60% were CD14+, i.e. 600 - fold increase in the number of CD14+ cells. The presence of two subpopulations of monocytes: CD14+CD16- and CD14++CD16+, which occurred at the ratio of about 2:1 was observed. They are clearly distinct from known main subpopulations of blood monocytes CD14+CD16+ and CD14++. Supplementation of cultures with VD3 increased the percentage of CD14+CD16- and decreased the proportion of CD14++CD16+. Monocytes/macrophages, mainly CD14++/16+ subpopulation secreted IL-12, but not TNF and IL-10, induced allo-MLR (mainly CD14++/16+ subpopulation), fagocytosed S.aureus (mainly CD14+/16- subpopulation), but did not produce intracellular O2-. Hence, this study shows that it is possible to generate from CD34+ haematopoietic progenitors two subpopulations of human monocytes. They are clearly distinct from those present in the peripheral blood im ; plicating the existence of novel monocyte subsets with distinct biological function that represent different stages in monocyte differentiation.

Place of publishing:

Kraków

Level of degree:

2 - studia doktoranckie

Degree discipline:

hematologia

Degree grantor:

Wydział Lekarski

Promoter:

Zembala, Marek

Date:

2007

Date issued:

2006

Type:

Praca doktorska

Call number:

ZB-105505

Language:

pol

Access rights:

nieograniczony

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