Isolation and culture of wapiti (Cervus elaphus) satellite cells

2000 ◽  
Vol 80 (2) ◽  
pp. 303-309
Author(s):  
N.M. Burton ◽  
L. Shipley ◽  
K. M. Byrne ◽  
J. L. Vierck ◽  
M. V. Dodson
Keyword(s):  
Cell Viability ◽  
Satellite Cells ◽  
Cervus Elaphus ◽  
Horse Serum ◽  
Media Effect ◽  
In Vitro Proliferation ◽  
Pig Skin ◽  
Defined Media ◽  
Proliferation And Differentiation

Myogenic satellite cells (SC) were isolated from the sternomandibularis muscles of two, 227 kg, male wapiti (Cervus elaphus) and studied in primary cell culture. Wapiti-derived SC were capable of attaching to culture substrata and following the myogenic program of proliferation and differentiation to form multinucleated myotubes. Wapiti SC attached equally well to pig skin gelatin (PSG), fibronectin (FN), Matrigel® and plastic (P > 00.05), but cell viability measured at 120 h varied depending on initial substratum type. Pig skin gelatin (0.02% wt vol−1) was chosen for the majority of subsequent experimentation for cost efficiency. The greatest amount of wapiti SC proliferation was observed in media containing 10% (vol/vol−1) horse serum (HS), 15% HS and 15% fetal bovine serum (FBS) (P > 0.05). Wapiti SC proliferated more when exposed to HS and FBS than to sheep serum (SS) (P < 0.05). No proliferation, differentiation or decrease in cell viability was observed in Dulbecco's Modified Eagle Medium (DMEM) + 1% HS, DMEM + 2% HS, DMEM + 3% HS or DMEM + 4% HS (P > 0.05) after 120 h in vitro. Proliferation of SC was doubled when insulin was added to both 10% HS- and 2% HS-containing media (P < 0.05). Although insulin alone in serum-containing media did not promote fusion of wapiti SC, two defined media (ITT and ITT-CF) that contain insulin did promote fusion of wapiti SC cultures. ITT-CF induced 3% fusion of wapiti SC into myotubes, and ITT induced 1% (P < 0.05). There was also an increase in total cell numbers in SC exposed to ITT-CF in comparison with ITT, ovine defined media (ODM) or ovine defined media-Modified (ODM-Mod))(P < 0.05). Although defined media differed in their ability to induce proliferation or differentiation (P < 0.05), the substrata on which the SC were plated did not influence the defined media effect on SC activity (P > 0.05). Satellite cells exposed to ITT and ITT-CF differed morphologically from SC exposed to ODM and ODM-Mod, which may suggest that formulation differences are influencing wapiti-derived SC proliferation and differentiation. Key words: Wapiti, satellite cells, primary culture, myotubes

In Vitro Proliferation and Differentiation of Human Hair Follicle Stem Cells on Chitosan-Skin Regenerating Template

2015 ◽  
Vol 05 (999) ◽  
pp. 1-1
Author(s):  
Abu Bakar Mohd Hilmi ◽  
Mohd Noor Norhayati ◽  
Ahmad Sukari Halim ◽  
Chin Keong Lim ◽  
Zulkifli Mustafa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hair Follicle ◽  
Human Hair ◽  
Human Hair Follicle ◽  
In Vitro Proliferation ◽  
Proliferation And Differentiation ◽  
Hair Follicle Stem Cells ◽  
Follicle Stem Cells

Sex, fiber-type, and age dependent in vitro proliferation of mouse muscle satellite cells

2011 ◽  
Vol 112 (10) ◽  
pp. 2825-2836 ◽  
Author(s):  
Raquel Manzano ◽  
Janne M. Toivonen ◽  
Ana C. Calvo ◽  
Francisco J. Miana-Mena ◽  
Pilar Zaragoza ◽  
...  
Keyword(s):  
Satellite Cells ◽  
Fiber Type ◽  
In Vitro Proliferation ◽  
Mouse Muscle ◽  
Muscle Satellite Cells ◽  
Age Dependent

scholarly journals miR-194-5p negatively regulates the proliferation and differentiation of rabbit skeletal muscle satellite cells

2020 ◽  
Author(s):  
Yu Shi ◽  
Xudong Mao ◽  
Mingcheng Cai ◽  
Shenqiang Hu ◽  
Xiulan Lai ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Satellite Cells ◽  
Mammalian Species ◽  
Muscle Growth ◽  
Luciferase Reporter ◽  
Stem Cell Population ◽  
Muscle Satellite Cells ◽  
Skeletal Muscle Satellite Cells ◽  
Proliferation And Differentiation

Abstract Skeletal muscle satellite cells (SMSCs), also known as a multipotential stem cell population, play a crucial role during muscle growth and regeneration. In recent years, numerous miRNAs have been associated with the proliferation and differentiation of SMSCs in a number of mammalian species; however, the regulatory mechanisms of miR-194-5p in rabbit SMSCs still remain scarce. In this study, miR-194-5p was first observed to be highly expressed in the rabbit leg muscle. Furthermore, both the mimics and inhibitor of miR-194-5p were used to explore its role in the proliferation and differentiation of rabbit SMSCs cultured in vitro. Results from both EdU and CCK8 assays showed that miR-194-5p inhibited the proliferation of SMSCs. Meanwhile, Mef2c was identified as a target gene of miR-194-5p based on the dual-luciferase reporter assay results. In addition, upregulation of miR-194-5p decreased the expression levels of Mef2c and MyoG during rabbit SMSCs differentiation on Days 3 and 7 of in vitro culture. Taken together, these data demonstrated that miR-194-5p negatively regulates the proliferation and differentiation of rabbit SMSCs by targeting Mef2c.

scholarly journals Existence of a pool of T-lymphocyte colony-forming cells (T-CFC) in human bone marrow and their place in the differentiation of the T- lymphocyte lineage

Blood ◽  
1981 ◽  
Vol 58 (5) ◽  
pp. 911-915 ◽  
Author(s):  
F Triebel ◽  
WA Robinson ◽  
AR Hayward ◽  
PG Goube de Laforest
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
T Lymphocyte ◽  
Cell Lineage ◽  
Marrow Graft ◽  
In Vitro Proliferation ◽  
Complement Dependent Cytotoxicity ◽  
Self Tolerance ◽  
Proliferation And Differentiation

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.

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

Blood ◽  
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

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.

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

Blood ◽  
2002 ◽  
Vol 99 (5) ◽  
pp. 1594-1601 ◽  
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.

scholarly journals Altered in vitro Proliferation of Mouse SOD1-G93A Skeletal Muscle Satellite Cells

2012 ◽  
Vol 11 (3) ◽  
pp. 153-164 ◽  
Author(s):  
Raquel Manzano ◽  
Janne M. Toivonen ◽  
Ana C. Calvo ◽  
Sara Oliván ◽  
Pilar Zaragoza ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Satellite Cells ◽  
In Vitro Proliferation ◽  
Muscle Satellite Cells ◽  
Skeletal Muscle Satellite Cells
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