1,25-dihydroxyvitamin D3 differentiates normal neutrophilic promyelocytes to monocytes/macrophages in vitro

• Published in: Blood Vol. 87; no. 7; pp. 2693 - 2701
• Main Authors: NAKAMURA, K, TAKAHASHI, T, SASAKI, Y, TSUYUOKA, R, OKUNO, Y, KURINO, M, OHMORI, K, IHO, S, NAKAO, K
• Format: Journal Article
• Language: English
• Published: Washington, DC The Americain Society of Hematology 01-04-1996
• Subjects: Biological and medical sciences; Calcitriol - pharmacology; Cell Differentiation - drug effects; Cell differentiation, maturation, development, hematopoiesis; Cell Lineage; Cell physiology; Cells, Cultured; Fundamental and applied biological sciences. Psychology; Granulocyte Colony-Stimulating Factor - pharmacology; Hematopoietic Stem Cells - cytology; Humans; Macrophages - cytology; Molecular and cellular biology; Monocytes - cytology; Neutrophils - cytology; Vitamin D; Monocyte; Cholecalciferol(1,25-dihydroxy); Promyelocyte; Neutrophil; Cell differentiation; Myeloid cell; In vitro; Precursor; Biological activity; Macrophage
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v87.7.2693.bloodjournal8772693

Although it is well established that the addition of 1,25-dihydroxyvitamin D3 (D3) to the culture of normal human granulocyte/macrophage progenitors induces monocyte/macrophage (Mo/M phi) colonies, the target cells of D3 in the Mo/M phi differentiation have not been identified. We examined whether neutrophilic promyelocytes are the target cells. As a source of the promyelocyte fraction, we used colonies after 5 days of culture (5-day colonies) of colony-forming unit-granulocyte. The culture contained granulocyte colony-stimulating factor (G-CSF) as the growth factor and generated only neutrophilic colonies. The promyelocytic nature of the 5-day colonies was confirmed morphologically, cytochemically, and ultrastructurally. After morphological evaluation on part of the individual colonies, they were transferred into new semisolid cultures with or without D3 (10(-7) mol/L) in the presence of G-CSF, then incubated for the subsequent 7 days. With D3, the colonies were loose, and all the constituent cells were morphologically small macrophages, which were positive for alpha-naphthyl butyrate (alpha NB) esterase, strongly positive for CD14 antigen, and plastic-adherent. While without D3, the colonies were rather compact, and all the constituent cells were morphologically mature neutrophils, which were positive for naphthol ASD-chloroacetate esterase and weakly positive for CD14 antigen. Secondary culture of the 8- or 10-day colonies with D3 induced a lower number of alpha NB-positive cells, in proportion to the percentage of promyelocytes at the time of transfer in each colony. Four days of secondary culture with D3 was sufficient to induce alpha NB-positive cells. G-CSF was not an essential factor to induce alpha NB-positive cells. These findings indicate that D3 differentiates normal human neutrophilic promyelocytes into the Mo/M phi lineage in vitro.