scholarly journals The role of insulin in transdifferentiated hepatocyte proliferation and function in serum‐free medium

2019 ◽  
Vol 23 (6) ◽  
pp. 4165-4178 ◽  
Author(s):  
Ce Gu ◽  
Panpan Li ◽  
Wei Liu ◽  
Yan Zhou ◽  
Wen‐Song Tan
Keyword(s):  
Hepatocyte Proliferation ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
And Function

scholarly journals A GPI-Anchored Protein, CD109, Protects Hematopoietic Progenitor Cells from Erythroid Differentiation Induced By TGF-β

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 3736-3736
Author(s):  
Tanabe Mikoto ◽  
Nguyen Hoang Maianh ◽  
Kohei Hosokawa ◽  
Noriharu Nakagawa ◽  
Luis Espinoza ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Erythroid Differentiation ◽  
Leukemia Cell Line ◽  
Serum Free Medium ◽  
Free Medium ◽  
Hematopoietic Stem ◽  
Serum Free ◽  
Company Limited ◽  
Immune Mediated

[Background] Glycosylphosphatidylinositol-anchored proteins (GPI-APs) on hematopoietic stem progenitor cells (HSPCs) may have some roles in the negative regulation of the HSPC commitment induced by inflammatory cytokines given the fact that progenies of GPI(-) HSPC are often detected in patients with immune-mediated bone marrow (BM) failure. CD109, one of the GPI-APs expressed by keratinocytes and HSPCs in humans, serves as a TGF-β co-receptor and is reported to inhibit TGF-β signaling in keratinocytes; however, the role of CD109 on HSPCs remains unknown. We previously demonstrated that TGF-β induced erythroid differentiation of TF-1 cells, a myeloid leukemia cell line that expresses CD109, in a dose-dependent manner and that knockout of the CD109 gene resulted in erythroid differentiation of TF-1 cells cultured in fetal bovine serum-containing medium, suggesting an inhibitory role of CD109 in the erythroid differentiation of HSPCs induced by low levels of TGF-β (Blood, 2018. 132 (Suppl.1) :3874). However, as most CD109 KO TF-1 cells changed into erythroid cells, they were unsuitable for investigating the role of CD109 in the erythroid differentiation induced by TGF-β. To overcome this issue, we prepared TF-1 cells and cord blood (CB) HSPCs in which the CD109 expression was transiently downregulated, and attempted to further clarify the role of CD109. [Methods] TF-1 cells and CD34+ cells isolated from CB mononuclear cells were treated with siRNA that was complementary to CD109 mRNA. CD109 knockdown cells were cultured for 4 days in serum-free medium supplemented with stem cell factor, thrombopoietin, and erythropoietin with or without TGF-β. In separate experiments, TF-1 cells were treated with phosphatidylinositol-specific phospholipase C (PIPL-C) treatment for 1 hour and were incubated in the presence or absence of TGF-β. CD109 KO TF-1 cells were incubated in serum-free medium (StemPro-34 SFM) for 14 days and their phenotype was determined using flow cytometry (FCM). The erythroid differentiation of the cells was assessed by testing the expression of glycophorin A (GPA) and iron staining. [Results] The down-regulation of CD109 in TF-1 cells by the siRNA treatment increased GPA expression in response to 12 ng/ml of TGF-β from 1.77% to 35.6%. The transient depletion of GPI-APs by PIPL-C also augmented the GPA expression induced by TGF-β from 1.27% to 6.77%. In both BM of healthy individuals and CB, CD109 was more abundantly expressed in Lin-CD34+CD38-CD90+CD45RA- hematopoietic stem cells (HSCs) than in Lin-CD34+CD38-CD90-CD45RA- multipotent progenitors (MPPs) and Lin-CD34+CD38+ HSPCs (Fig. 1). The treatment of CB cells with siRNA reduced the CD109 expression in Lin-CD34+CD38+ cells from 55.9% to 23.1%. TGF-β induced the expression of GPA in Lin-CD34+CD38+CD123-CD45RA- megakaryocyte-erythrocyte progenitor cells (MEPs) of CD109 knockdown cells to a greater degree than the control counterpart (Fig. 2). During 14-day serum-free culture, GPA-positive CD109 KO TF-1 cells died, and similarly to WT TF-1 cells, most surviving CD109 KO TF-1 cells were GPA-negative. TGF-β treatment induced erythroid differentiation in CD109 KO TF-1 cells to a greater degree than in WT TF-1 cells. [Conclusions] CD109 plays a key role in the inhibition of TF-1 erythroid differentiation in response to TGF-β. CD109 may suppress TGF-β signaling, and the lack of CD109 may make PIGA-mutated HSPCs more sensitive to TGF-β, thus leading to the preferential commitment of the mutant erythroid progenitor cells to mature red blood cells in immune-mediated BM failure. Disclosures Yamazaki: Novartis Pharma K.K.: Honoraria; Sanofi K.K.: Honoraria; Nippon Shinyaku Co., Ltd.: Honoraria. Nakao:Novartis Pharma K.K: Honoraria; Bristol-Myers Squibb: Honoraria; Takeda Pharmaceutical Company Limited: Honoraria; Celgene: Honoraria; Ono Pharmaceutical: Honoraria; Chugai Pharmaceutical Co.,Ltd: Honoraria; Kyowa Kirin: Honoraria; Alaxion Pharmaceuticals: Honoraria; Ohtsuka Pharmaceutical: Honoraria; Daiichi-Sankyo Company, Limited: Honoraria; Janssen Pharmaceutical K.K.: Honoraria; SynBio Pharmaceuticals: Consultancy.

Possible role of neuropeptide degrading enzymes on thyroliberin secretion in fetal hypothalamic cultures grown in serum free medium

Neuropeptides ◽  
1986 ◽  
Vol 7 (2) ◽  
pp. 125-138 ◽  
Author(s):  
Annie Faivre-Bauman ◽  
Catherine Loudes ◽  
Alain Barret ◽  
Andrée Tixier-Vidal ◽  
Karl Bauer
Keyword(s):  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Degrading Enzymes

Role of contraction in the structure and growth of neonatal rat cardiocytes

1987 ◽  
Vol 253 (6) ◽  
pp. H1391-H1399 ◽  
Author(s):  
T. A. Marino ◽  
L. Kuseryk ◽  
I. K. Lauva
Keyword(s):  
Fine Structure ◽  
Surface Area ◽  
Neonatal Rat ◽  
Serum Free Medium ◽  
Free Medium ◽  
Growth Promoters ◽  
Volume Number ◽  
Serum Free ◽  
Substantial Growth

The aim of this study was to determine the role of contraction in the regulation of neonatal rat cardiocyte growth in size. To accomplish this objective, experiments were done on four groups of cardiocytes: 1) quiescent cardiocytes attached to a substrate, 2) contracting cardiocytes attached to a substrate, 3) quiescent cardiocytes not attached to a substrate, adn 4) contracting cardiocytes not attached to a substrate. The cardiocytes were grown in both serum-free and serum-supplemented media for up to 1 wk, and cardiocyte surface area, volume, number, and fine structure were evaluated. The most important result of this study was that cardiocytes, which are attached to a substrate and stimulated to contract, grow in size. However, neither contraction alone nor attachment to a substrate by itself resulted in neonatal cardiocyte growth in size in defined serum-free medium. Another important finding was that other nonspecific growth promoters, such as those found in serum, stimulated more substantial growth in cardiocyte size in contracting cardiocytes that were attached to a substrate.

Role of iron in transferrin-dependent lymphocyte mitogenesis in serum-free medium

1982 ◽  
Vol 139 (2) ◽  
pp. 419-422 ◽  
Author(s):  
A NOVOGRODSKY ◽  
A RAVID ◽  
T GLASER ◽  
A RUBIN ◽  
K STENZEL
Keyword(s):  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Lymphocyte Mitogenesis

The Role of Endogenous Stromal Cell-Derived Factor-1 in the Survival and Growth of Acute Myeloid Leukemia Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1502-1502
Author(s):  
Ha-Yon Kim ◽  
Yoon-Suk Oh ◽  
Ik-Chan Song ◽  
Hyo-Jin Lee ◽  
Hwan-Jung Yun ◽  
...  
Keyword(s):  
Cell Lines ◽  
Myeloid Leukemia ◽  
Leukemia Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Knock Down ◽  
Cd34 Cells ◽  
Stromal Cell Derived Factor

Abstract Abstract 1502 Stromal cell-derived factor-1 (SDF-1) is constitutively expressed and produced by stromal cells in the bone marrow (BM). The protein exerts its functions by binding to its receptor CXCR4, thus guiding the homing of hematopoietic stem/progenitor cells and leukemia cells to the BM. CD34+ cells purified from normal adult peripheral blood have been shown to express and produce SDF-1, which exerted an antiapoptotic effect on CD34+ cell through an autocrine/paracrine loop. Acute myeloid leukemia (AML) cells have also been shown to express and produce SDF-1, but the role of SDF-1 in this context is largely unknown. Thus, the aim of the present study was to determine the function of SDF-1 produced by AML cells. Using siRNA technology, SDF-1 was knocked-down in AML cells and subsequent biological alterations occurring in the cells were evaluated in vitro. All AML cell lines used in this study (U937, K562, KG1a, HL-60 and MO7e) as well as primary CD34+ cells obtained from the BM of 5 patients with AML expressed SDF-1 mRNA at various levels, as revealed by semi-quantitative and real time RT-PCR analysis. SDF-1 was detected by Western blotting analysis and ELISA allowed for the detection of SDF-1 in the supernatants of cultures of AML cells grown for 3 days in a serum-free medium, indicating that AML cells, similar to normal CD34+ cells, are able to produce and secrete SDF-1. Exogenous SDF-1 (added at a concentration of up to 200 ng/mL) alone had no discernible effects on the survival or proliferation of either the cell lines or the primary CD34+ AML cells maintained in a serum-free medium, but was able to induce the internalization of CXCR4 and CXCR7. Transfection of five AML cell lines with SDF-1 siRNA diminished the SDF-1 mRNA levels by up to 90%. The knock-down cells were stable for at least 3 days. SDF-1 knock-down did not affect the serum deprivation-induced apoptosis of the cells. Cell surface expression of CXCR4 was not changed, whereas cytoplasmic CXCR4 was significantly upregulated along with the upregulation of SDF-1 mRNA. Neither cell surface nor cytoplasmic expression of CXCR7 was affected by the knock-down of SDF-1. Conversely, the knock-down of CXCR7, but not CXCR4, upregulated the SDF-1 mRNA expression in the transfected cells. No changes in the transmembrane and trnasendothelial migration of the knock-down cells were observed. In contrast, the knock-down of SDF-1 inhibited the spontaneous proliferation of the U937, K562, and KG1a cells by 20–30% during a 3-day incubation in a serum-free medium. These results indicate that endogenous SDF-1 of AML cells functions as an autocrine growth factor. Disclosures: No relevant conflicts of interest to declare.

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