In vitro proliferation and differentiation of hepatic oval cells and their potential capacity for intrahepatic transplantation

Abstract The existence and characteristics of bone marrow T-cell progenitors have not yet been established in man. Several pieces of evidence such as the reconstitution of certain immunodeficiencies by bone marrow graft suggest that T-cell precursors are present in the bone marrow. We report the growth of T-cell colonies from bone marrow populations using PHA-stimulated lymphocyte-conditioned medium containing T-cell growth factor (TCGF). Rosetting experiments and complement-dependent cytotoxicity assays with monoclonal antibodies indicate that the bone marrow T colony-forming cells (T-CFC) are E- OKT 3- and la+, i.e., immature progenitors. The colonies derived from these cells have the phenotype of mature T cells: E + OKT 3 + la- with either helper (OKT 4+) and suppressor (OKT 8 +) antigens. These results suggest that a thymic microenvironment may not be necessary for the in vitro proliferation and differentiation of the T-cell lineage in adult humans. These methodologies may permit direct investigation of early phenomena concerning the T-cell lineage, such as the acquisition of self-tolerance, the formation of a repertoire of specificities, and the HLA restriction phenomena that we believe takes place before the thymic maturation.

Download Full-text

In vitro proliferation and differentiation of canine bone marrow derived mesenchymal stem cells over hydroxyl functionalized CNT substrates

Biotechnology Reports ◽

10.1016/j.btre.2019.e00387 ◽

2019 ◽

Vol 24 ◽

pp. e00387

Author(s):

A.P. Madhusoodan ◽

Kinsuk Das ◽

Bhabesh Mili ◽

Kuldeep Kumar ◽

Ajay Kumar ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

In Vitro Proliferation ◽

Proliferation And Differentiation

Download Full-text

Common and pre-B acute lymphoblastic leukemia cells express interleukin 2 receptors, and interleukin 2 stimulates in vitro colony formation

Blood ◽

10.1182/blood.v66.3.556.556 ◽

1985 ◽

Vol 66 (3) ◽

pp. 556-561 ◽

Cited By ~ 44

Author(s):

I Touw ◽

R Delwel ◽

R Bolhuis ◽

G van Zanen ◽

B Lowenberg

Keyword(s):

Acute Lymphoblastic Leukemia ◽

Colony Formation ◽

B Lymphocyte ◽

Lymphoblastic Leukemia ◽

Interleukin 2 ◽

Colony Growth ◽

Leukemic Cells ◽

In Vitro Proliferation ◽

Proliferation And Differentiation

Abstract The role of interleukin 2 (IL 2) as a possible regulator of in vitro proliferation and differentiation of non-T acute lymphoblastic leukemia (ALL) cells was investigated. For this purpose, leukemic cells from the blood or bone marrow of eight untreated patients with common or pre-B ALL were analyzed using the anti-Tac monoclonal antibody (reactive with the IL 2 receptor) in indirect immunofluorescence. The receptors for IL 2, which were initially absent from the cell surface, were induced on high percentages of the ALL cells after the in vitro exposure to the lectin phytohemagglutinin or the phorbol ester 12-O- tetradecanoylphorbol-13-acetate in six patients, suggesting that the cells had become sensitive to IL 2. In colony cultures to which feeder leukocytes and IL 2 had been added, colony growth was obtained in five of eight cases. Whereas the cells from one patient formed colonies in the absence of exogenous stimuli, the cells from others were dependent on the addition of feeder leukocytes plus IL 2. In the latter cases, feeder leukocytes alone, releasing some IL 2, stimulated growth suboptimally at different cell concentrations. Their stimulative effect was significantly enhanced when leukocyte-derived IL 2 or pure recombinant IL 2 was supplemented. Alone, IL 2 (up to 500 U/mL) did not support colony formation. Apparently, IL 2 and feeder leukocytes are both required for the induction of colonies in these cases of ALL. From cell sorting of fluorescent anti-common ALL antigen (CALLA) stained cells it appeared that colonies descended from cells with high as well as low or negative CALLA expression. Immunophenotyping demonstrated the presence of the original leukemia markers on colony cells, but was not indicative of maturation of ALL toward more differentiated B cells. We suggest that IL 2 can stimulate the in vitro proliferation of certain neoplastic B lymphocyte progenitors.

Download Full-text

In vitro proliferation and differentiation of erythroid progenitors from patients with myelodysplastic syndromes: evidence for Fas-dependent apoptosis

Blood ◽

10.1182/blood.v99.5.1594 ◽

2002 ◽

Vol 99 (5) ◽

pp. 1594-1601 ◽

Cited By ~ 94

Author(s):

Yann-Erick Claessens ◽

Didier Bouscary ◽

Jean-Michel Dupont ◽

Françoise Picard ◽

Josiane Melle ◽

...

Keyword(s):

Myelodysplastic Syndromes ◽

Liquid Culture ◽

Fas Ligand ◽

Chimeric Protein ◽

Refractory Anemia ◽

Erythroid Progenitors ◽

In Vitro Proliferation ◽

Cd34 Cells ◽

Proliferation And Differentiation

Erythropoiesis results from the proliferation and differentiation of pluripotent stem cells into immature erythroid progenitors (ie, erythroid burst-forming units (BFU-Es), whose growth, survival, and terminal differentiation depends on erythropoietin (Epo). Ineffective erythropoiesis is a common feature of myelodysplastic syndromes (MDS). We used a 2-step liquid-culture procedure to study erythropoiesis in MDS. CD34+ cells from the marrow of patients with MDS were cultured for 10 days in serum-containing medium with Epo, stem cell factor, insulinlike growth factor 1, and steroid hormones until they reached the proerythroblast stage. The cells were then placed in medium containing Epo and insulin for terminal erythroid differentiation. Numbers of both MDS and normal control cells increased 103fold by day 15. However, in semisolid culture, cells from patients with refractory anemia (RA) with ringed sideroblasts and RA or RA with excess of blasts produced significantly fewer BFU-Es than cells from controls. Fluorescence in situ hybridization analysis of interphase nuclei from patients with chromosomal defects indicated that abnormal clones were expanded in vitro. Epo-signaling pathways (STAT5, Akt, and ERK 1/2) were normally activated in MDS erythroid progenitors. In contrast, apoptosis was significantly increased in MDS cells once they differentiated, whereas it remained low in normal cells. Fas was overexpressed on freshly isolated MDS CD34+ cells and on MDS erythroid cells throughout the culture. Apoptosis coincided with overproduction of Fas ligand during the differentiation stage and was inhibited by Fas-Fc chimeric protein. Thus, MDS CD34+-derived erythroid progenitors proliferated normally in our 2-step liquid culture with Epo but underwent abnormal Fas-dependent apoptosis during differentiation that could be responsible for the impaired erythropoiesis.

Download Full-text

In vitro and in vivo characteristics of hepatic oval cells modified with human hepatocyte growth factor

Cellular & Molecular Biology Letters ◽

10.2478/s11658-013-0104-1 ◽

2013 ◽

Vol 18 (4) ◽

Cited By ~ 2

Author(s):

Zhu Li ◽

Juan Chen ◽

Li Li ◽

Jiang-Hua Ran ◽

Xue-Hua Li ◽

...

Keyword(s):

Liver Transplantation ◽

Growth Factor ◽

Liver Function ◽

Hepatocyte Growth Factor ◽

Orthotopic Liver Transplantation ◽

Peak Time ◽

Oval Cells ◽

Hepatic Oval Cells ◽

Hepatocyte Growth

AbstractHepatocyte growth factor (HGF) is a multifunctional growth factor that controls cell scattering. It has been suggested that it regulates the proliferation of hepatic oval cells (HOCs). Using a HOC line that stably expresses the human HGF gene (hHGF), we investigated the in vitro proliferation and differentiation characteristics of hHGF-modified HOCs and explored their potential capacity for intrahepatic transplantation. A modified 2-acetylaminofluorene and partial hepatectomy (2-AAF/PH) model was established to activate the proliferation of oval cells in the rat liver. HOCs were transfected with the pBLAST2-hHGF plasmid and hHGF-carrying HOCs were selected based on blasticidin resistance. The level of hHGF secretion was determined via ELISA. Cell proliferation was determined using the MTT assay. Differentiation was induced by growth factor withdrawal. A two-cuff technique was used for orthotopic liver transplantation, and HOCs or hHGF-modified HOCs were transplanted into the recipients. The levels of biochemical indicators of liver function were measured after transplantation. An HOC line stably expressing hHGF was established. The transfected line showed greater hHGF secretion than normal HOCs. The hHGF gene promoted the proliferation capability of HOCs by reducing the peak time in vitro. The hHGF-modified HOCs differentiated into hepatocytes and bile duct epithelial cells upon growth factor withdrawal in vitro. In addition, hHGF-modified HOC transplantation significantly prolonged the median survival time (MST) and improved the liver function of recipients compared to HOC transplant recipients and nontransplanted controls. Our results indicate that hHGF-modified HOCs may have valuable properties for therapeutic liver regeneration after orthotopic liver transplantation.

Download Full-text

128. Transduction of Hepatic Oval Cells, Stem Cells, In Vitro by Adeno-Associated Virus Vectors

Molecular Therapy ◽

10.1016/j.ymthe.2005.06.133 ◽

2005 ◽

Vol 11 ◽

pp. S52

Keyword(s):

Stem Cells ◽

Oval Cells ◽

Adeno Associated Virus ◽

Virus Vectors ◽

Hepatic Oval Cells

Download Full-text

The role of hormones and prostanoids in the in vitro proliferation and differentiation of human myoblasts

Experimental Cell Research ◽

10.1016/0014-4827(87)90386-7 ◽

1987 ◽

Vol 172 (2) ◽

pp. 265-281 ◽

Cited By ~ 47

Author(s):

Rosalind J. Zalin

Keyword(s):

In Vitro Proliferation ◽

Proliferation And Differentiation

Download Full-text

Common and pre-B acute lymphoblastic leukemia cells express interleukin 2 receptors, and interleukin 2 stimulates in vitro colony formation

Blood ◽

10.1182/blood.v66.3.556.bloodjournal663556 ◽

1985 ◽

Vol 66 (3) ◽

pp. 556-561

Author(s):

I Touw ◽

R Delwel ◽

R Bolhuis ◽

G van Zanen ◽

B Lowenberg

Keyword(s):

Acute Lymphoblastic Leukemia ◽

Colony Formation ◽

B Lymphocyte ◽

Lymphoblastic Leukemia ◽

Interleukin 2 ◽

Colony Growth ◽

Leukemic Cells ◽

In Vitro Proliferation ◽

Proliferation And Differentiation

The role of interleukin 2 (IL 2) as a possible regulator of in vitro proliferation and differentiation of non-T acute lymphoblastic leukemia (ALL) cells was investigated. For this purpose, leukemic cells from the blood or bone marrow of eight untreated patients with common or pre-B ALL were analyzed using the anti-Tac monoclonal antibody (reactive with the IL 2 receptor) in indirect immunofluorescence. The receptors for IL 2, which were initially absent from the cell surface, were induced on high percentages of the ALL cells after the in vitro exposure to the lectin phytohemagglutinin or the phorbol ester 12-O- tetradecanoylphorbol-13-acetate in six patients, suggesting that the cells had become sensitive to IL 2. In colony cultures to which feeder leukocytes and IL 2 had been added, colony growth was obtained in five of eight cases. Whereas the cells from one patient formed colonies in the absence of exogenous stimuli, the cells from others were dependent on the addition of feeder leukocytes plus IL 2. In the latter cases, feeder leukocytes alone, releasing some IL 2, stimulated growth suboptimally at different cell concentrations. Their stimulative effect was significantly enhanced when leukocyte-derived IL 2 or pure recombinant IL 2 was supplemented. Alone, IL 2 (up to 500 U/mL) did not support colony formation. Apparently, IL 2 and feeder leukocytes are both required for the induction of colonies in these cases of ALL. From cell sorting of fluorescent anti-common ALL antigen (CALLA) stained cells it appeared that colonies descended from cells with high as well as low or negative CALLA expression. Immunophenotyping demonstrated the presence of the original leukemia markers on colony cells, but was not indicative of maturation of ALL toward more differentiated B cells. We suggest that IL 2 can stimulate the in vitro proliferation of certain neoplastic B lymphocyte progenitors.

Download Full-text