scholarly journals Quantitation of human in vitro megakaryocytopoiesis by radioimmunoassay

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1334-1339 ◽  
Author(s):  
BW Grant ◽  
WL Nichols ◽  
LA Solberg ◽  
DJ Yachimiak ◽  
KG Mann
Keyword(s):  
Monoclonal Antibody ◽  
Growth Factors ◽  
Liquid Culture ◽  
Sensitive Assay ◽  
Semisolid Medium ◽  
Platelet Production ◽  
Isolation And Characterization ◽  
Marrow Cultures

Abstract The isolation and characterization of human megakaryocyte growth factors has been hampered because evaluation of megakaryocyte growth in semisolid medium requires both lengthy incubation and visual quantitation. In addition, colony formation requires cell division, while most regulation of platelet production may involve individual, nonproliferating differentiating megakaryocytes. We have developed a radioimmunoassay (RIA) that makes use of an iodinated murine monoclonal antibody (MoAb) specific for platelet/megakaryocyte glycoprotein IIb/IIIa (GPIIb/IIIa) to measure megakaryocyte production in liquid marrow culture. This assay is sensitive to 3 X 10(3) platelets (roughly 30 megakaryocytes) and linear up to 1 X 10(6) platelets, and thus it provides a useful range for quantitating megakaryocyte production in in vitro marrow culture. Significant differences (threefold to fivefold) in megakaryocyte/platelet-specific GPIIb/IIIa complex are detected between stimulated and unstimulated marrow cultures by day 7, although antigen accrual in stimulated cultures continues through at least day 16. Conditions that promote megakaryocyte growth in semisolid medium (ie, aplastic plasma and PHA-LCM) have also been facilitory in liquid culture. This rapid and sensitive assay for cell-bound GPIIb/IIIa should facilitate recognition and isolation of megakaryocyte and platelet growth factors.

scholarly journals Quantitation of human in vitro megakaryocytopoiesis by radioimmunoassay

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1334-1339
Author(s):  
BW Grant ◽  
WL Nichols ◽  
LA Solberg ◽  
DJ Yachimiak ◽  
KG Mann
Keyword(s):  
Monoclonal Antibody ◽  
Growth Factors ◽  
Liquid Culture ◽  
Sensitive Assay ◽  
Semisolid Medium ◽  
Platelet Production ◽  
Isolation And Characterization ◽  
Marrow Cultures

The isolation and characterization of human megakaryocyte growth factors has been hampered because evaluation of megakaryocyte growth in semisolid medium requires both lengthy incubation and visual quantitation. In addition, colony formation requires cell division, while most regulation of platelet production may involve individual, nonproliferating differentiating megakaryocytes. We have developed a radioimmunoassay (RIA) that makes use of an iodinated murine monoclonal antibody (MoAb) specific for platelet/megakaryocyte glycoprotein IIb/IIIa (GPIIb/IIIa) to measure megakaryocyte production in liquid marrow culture. This assay is sensitive to 3 X 10(3) platelets (roughly 30 megakaryocytes) and linear up to 1 X 10(6) platelets, and thus it provides a useful range for quantitating megakaryocyte production in in vitro marrow culture. Significant differences (threefold to fivefold) in megakaryocyte/platelet-specific GPIIb/IIIa complex are detected between stimulated and unstimulated marrow cultures by day 7, although antigen accrual in stimulated cultures continues through at least day 16. Conditions that promote megakaryocyte growth in semisolid medium (ie, aplastic plasma and PHA-LCM) have also been facilitory in liquid culture. This rapid and sensitive assay for cell-bound GPIIb/IIIa should facilitate recognition and isolation of megakaryocyte and platelet growth factors.

Isolation and characterization of rabbit cardiac endothelial cells: Response to cyclic strain and growth factors in vitro

1992 ◽  
Vol 53 (4) ◽  
pp. 331-334 ◽  
Author(s):  
Mark D. Widmann ◽  
George V. Letsou ◽  
Stacie Phan ◽  
John C. Baldwin ◽  
Bauer E. Sumpio
Keyword(s):  
Endothelial Cells ◽  
Growth Factors ◽  
Cyclic Strain ◽  
Isolation And Characterization

PENGEMBANGAN TEKNOLOGI SELEKSI IN VITRO BERULANG (REPEAT CYCLING–IN VITRO SELECTION) TOLERAN STRES KEKERINGAN UNTUK PERBAIKAN MUTU GENETIK DAN PRODUKSI TANAMAN KEDELAI BERMIKORIZA

2017 ◽  
Vol 1 (1) ◽  
pp. 74-84
Author(s):  
Ahmad Riduan ◽  
Rainiyati Rainiyati ◽  
Yulia Alia
Keyword(s):  
Arbuscular Mycorrhizae ◽  
In Vitro Selection ◽  
Soil Samples ◽  
Single Spore ◽  
Isolation And Characterization ◽  
Single Spore Culture ◽  
Spore Culture ◽  
Glomus Sp

Every plant rhizospheres in any ecosystem there are various living microorganisms including Arbuscular Mycorrhizae Fungi (AMF).  An isolation and characterization is required to investigate the species or type of the AMF. This research was aimed at studying the isolation and characterization of AMF sporulation in soybean rhizospheres in Jambi Province. The results of evaluation on soil samples before trapping showed that there are spores from three genus of AMF twelve types Glomus , two types Acaulospora and one type of Enthrophospora.  Following single spore culture in soybean rhizosphere, 5 spore types were obtained:  Glomus sp-1, Glomus sp-4, Glomus sp-7, Glomus sp-8 Glomus sp-10.

Isolation and Characterization of a Monoclonal Antibody against Pig Kidney Sodium- and Potassium-Activated ATPase1

1985 ◽  
Vol 98 (1) ◽  
pp. 209-217 ◽  
Author(s):  
Osamu URAYAMA ◽  
Hideaki NAGAMUNE ◽  
Makoto NAKAO ◽  
Yukichi HARA ◽  
Hiroyuki SUGIYAMA ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Pig Kidney ◽  
Isolation And Characterization ◽  
Sodium And Potassium

Generation and characterization of a human monoclonal antibody that neutralizes diverse HIV-1 isolates in vitro

AIDS ◽  
1992 ◽  
Vol 6 (1) ◽  
pp. 17-24 ◽  
Author(s):  
Douglas F. Lake ◽  
Takashi Kawamura ◽  
Takami Tomiyama ◽  
W. Edward Robinson ◽  
Yoh-ichi Matsumoto ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Human Monoclonal Antibody ◽  
Hiv 1

Arginine synthesis by hepatomas in vitro. II. Isolation and characterization of Morris hepatoma variants unable to convert ornithine to arginine, and modulation of urea-cycle enzymes by dexamethasone and cyclic-AMP

1984 ◽  
Vol 68 (1) ◽  
pp. 305-319
Author(s):  
S.J. Goss
Keyword(s):  
Cyclic Amp ◽  
Liver Cells ◽  
Isolation And Characterization ◽  
Rat Liver Cells ◽  
Morris Hepatoma ◽  
Ornithine Transcarbamoylase ◽  
Cellular Incorporation

‘77orn’, a derivative of the Morris rat hepatoma 7777, stably expresses high levels of ornithine transcarbamoylase (OTC) and carbamoylphosphate synthetase I (CPS-I), and is able to grow indefinitely in ornithine-medium (medium with ornithine in place of arginine). Variants that have lost this ability are isolated from 77orn by a ‘suicide’ selective technique dependent on the cellular incorporation of [3H]ornithine. These variants, which have reduced levels of CPS-I, or of both CPS-I and OTC, are shown to have developed multiple hormonal requirements; their enzyme deficiencies can be reversed by use of an appropriately supplemented medium. In particular, CPS-I is inducible by dexamethasone and dibutyryl-cyclic-AMP in combination. Cholera toxin can be used instead of cyclic-AMP, but then butyrate is additionally required if the induction is to be maintained in the long term. The use of these agents in excess can depress OTC. Several other hepatomas, and alos explanted foetal rat liver cells, have similar requirements for CPS-I expression. It is argued that multiple hormonal requirements for CPS-I production are normal in liver cells in vitro, and that hormone-independent hepatomas should be regarded as abnormal. The implications of this for the somatic cell genetic investigation of differentiation are briefly discussed.

Isolation and characterization of phosphate solubilizing bacteria from the rhizospheric soil of Ponthenpuzha forest, Pathanamthitta (District), Kerala

2021 ◽  
Vol 16 (8) ◽  
pp. 110-117
Author(s):  
Kannan Abhirami ◽  
K. Jayakumar
Keyword(s):  
Bacterial Isolate ◽  
Phosphate Solubilizing Bacteria ◽  
Cymbopogon Citratus ◽  
16S Rrna Sequence ◽  
Isolation And Characterization ◽  
16S Rrna Sequence Analysis ◽  
Phosphate Solubilizing ◽  
Level Identification

Phosphorous is considered as a major parameter for crop yield. Its availability to plant is independent of its abundance. For the plants to utilize phosphorous, it is to be converted to absorbable form. Here, the part rendered by phosphate solubilizing bacteria is significant for it plays a crucial role in the formation of plant usable phosphate from organic forms. In the present work, an effort had been made to isolate and identify phosphate solubilising bacterial isolate from the rhizhospheric soils of various plants in Ponthenpuzha forest. One of the isolate from Cymbopogon citrates responded positively to Pikovskaya’s medium by producing a halo zone during in vitro culture. Colony features and 16S rRNA sequence analysis identified the isolate as Burkholderia sps. We have reported the presence of genus Burkholderia in the rhizospheric zone of Cymbopogon citratus. Further studies are warranted for species level identification of the isolate.

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