Ploidy instability in long-term in vitro cultures of Coffea arabica L. monitored by flow cytometry

2012 ◽  
Vol 68 (3) ◽  
pp. 533-538 ◽  
Author(s):  
Wellington Ronildo Clarindo ◽  
Carlos Roberto Carvalho ◽  
Maria Andréia Corrêa Mendonça
Keyword(s):  
Flow Cytometry ◽  
Coffea Arabica ◽  
In Vitro Cultures ◽  
Coffea Arabica L

scholarly journals Improved Conservation of Coffee (Coffea arabica L.) Germplasm via Micropropagation and Cryopreservation

Agronomy ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1861
Author(s):  
Yanelis Castilla Valdés ◽  
Mukund R. Shukla ◽  
María Esther González Vega ◽  
Praveen K. Saxena
Keyword(s):  
Coffea Arabica ◽  
Plant Genetic Resources ◽  
Ex Situ ◽  
Zygotic Embryos ◽  
Agricultural Crop ◽  
Temporary Immersion ◽  
Droplet Vitrification ◽  
Coffea Arabica L

Coffee (Coffea spp.) is an important tropical agricultural crop that has significant economic and social importance in the world. The ex situ conservation of plant genetic resources through seeds is not feasible due to the sensitivity of coffee seed to desiccation and low temperatures. The cryopreservation of zygotic embryos may allow for an efficient and long-term storage of coffee germplasm. This study describes the cryopreservation methods for conserving zygotic embryos of Coffea arabica L. for the long-term conservation of currently available germplasm. Zygotic embryos were successfully cryopreserved in liquid nitrogen at −196 °C under controlled environmental conditions with either droplet-vitrification or encapsulation–vitrification protocols without dehydration. Zygotic embryos had the highest regrowth (100%) following droplet-vitrification cryopreservation using the Plant Vitrification Solution 3 (PVS3) for 40 min at 23 °C. In the case of encapsulation–vitrification using PVS3 for 40 min at 23 °C, the embryo regeneration response was 78%. Plantlets were recovered following shoot multiplication using a temporary immersion system (TIS) and in vitro rooting. The prolific rooting of shoots was observed after 4 weeks of culture in the liquid medium with plugs made of the inert substrate Oasis® In vitro Express (IVE) compared to the semi-solid medium. The successful cryopreservation of coffee zygotic embryos using droplet vitrification and encapsulation–vitrification followed by micropropagation in temporary immersion culture system has not been reported earlier and together these technologies are anticipated to further facilitate the initiatives for the conservation and distribution of coffee germplasm.

scholarly journals Anatomy, Flow Cytometry, and X-Ray Tomography Reveal Tissue Organization and Ploidy Distribution in Long-Term In Vitro Cultures of Melocactus Species

2020 ◽  
Vol 11 ◽  
Author(s):  
Gabriela Torres-Silva ◽  
Elyabe Monteiro Matos ◽  
Ludmila Freitas Correia ◽  
Evandro Alexandre Fortini ◽  
Wellington Santos Soares ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
In Vitro Cultures ◽  
X Ray ◽  
Tissue Organization

71 TECHNIQUE FOR ESTABLISHMENT OF LONG TERM IN VITRO CULTURES OF PRIMARY MDS CELLS

2015 ◽  
Vol 39 ◽  
pp. S35
Author(s):  
C. Ichim ◽  
D. Koos ◽  
T. Ichim ◽  
R. Wells
Keyword(s):  
In Vitro Cultures

scholarly journals Corrigendum: Is 8% O2 more normoxic than 21% O2 for long-term in vitro cultures of human primary term cytotrophoblasts?

2018 ◽  
Vol 24 (5) ◽  
pp. 283-283
Author(s):  
C L Depoix ◽  
F Haegeman ◽  
F Debiève ◽  
C Hubinont
Keyword(s):  
In Vitro Cultures

scholarly journals Loss of primitive hematopoietic progenitors in patients with human immunodeficiency virus infection

Blood ◽  
1996 ◽  
Vol 88 (12) ◽  
pp. 4568-4578 ◽  
Author(s):  
A Marandin ◽  
A Katz ◽  
E Oksenhendler ◽  
M Tulliez ◽  
F Picard ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Human Immunodeficiency Virus ◽  
Long Term Culture ◽  
Cd34 Cells ◽  
Immunodeficiency Virus ◽  
Cell Fractions ◽  
Cd38 Antigen ◽  
Hiv 1

A number of hematologic abnormalities, including cytopenias, have been observed in patients with human immunodeficiency virus (HIV) infection. To elucidate their mechanisms, primitive cells from bone marrow aspirates of 21 patients with HIV-1 infection were quantitated by flow cytometry. The mean percentage of CD34+ cells is not significantly altered in HIV-1-infected patients in comparison with HIV-1- seronegative controls. In contrast, two- and three-color immunofluorescence analysis showed that in all HIV-1 samples, most CD34+ cells coexpressed the CD38 antigen. The proportion of HIV-1- derived CD34+ cells that did not express the CD38 antigen was significantly lower (HIV-1+: mean, 1.73%; controls: mean, 14%; P < .0005) than in controls. Moreover, of Thy-1+ cells, the proportion of CD34+ cells was twofold lower in HIV-1-infected patients (HIV-1+: mean, 12%; controls, 25%, P < .0005), which suggests that phenotypically primitive cells are depleted in HIV-1 infection. In vitro functional analysis in long-term cultures of sorted CD34+ cells from seven HIV-1 patients showed that CD34+ cells from HIV-1 patients generated much fewer colonies both in the nonadherent and adherent layers than CD34+ cells from controls after 5 weeks of culture (10-fold and four-fold less, respectively). Precise long-term culture initiating cell (LTC-IC) frequency in the CD34+ cell population was determined in three patients by limiting dilution and was markedly decreased in comparison to that of normal controls (from twofold to > sevenfold decreased). To determine if primitive cells were infected by HIV-1, both methylcellulose colonies generated from long-term culture of CD34+ cells and various CD34+ cell fractions purified by flow cytometry were evaluated for the presence of HIV-1 by polymerase chain reaction (PCR). Progeny from long-term culture was HIV-1-negative in three samples. In addition, using a sensitive PCR technique, the HIV-1 genome could not be detected in CD34+, CD34+/CD38-, and CD34+/CD4+ cells. These data show that hematologic disorders in HIV disease may be the consequence of a deficit of primitive cells. However, direct infection of these cells by HIV-1 does not seem to be responsible for this defect.

scholarly journals In vitro studies on lymphocyte differentiation. I. Long term in vitro culture of cells giving rise to functional lymphocytes in irradiated mice.

1978 ◽  
Vol 148 (3) ◽  
pp. 823-828 ◽  
Author(s):  
J W Schrader ◽  
S Schrader
Keyword(s):  
B Lymphocytes ◽  
Cultured Cells ◽  
Bone Marrow Cells ◽  
In Vitro Cultures ◽  
Mouse Bone Marrow ◽  
Graft Versus Host ◽  
Receptor Repertoire ◽  
Mouse Bone Marrow Cells

In vitro cultures of mouse bone marrow cells, maintained for periods up to 7 wk, were shown to contain cells able to repopulate irradiated hosts with T and B lymphocytes. The lymphocytes were fully functional and there did not appear to be any gross restriction of their receptor repertoire. The cultured cells reconstituted irradiated semiallogenic hosts without evidence of graft-versus-host disease, suggesting that culture of donor marrow might be a useful preliminary to transplantation when tissue matching is incomplete.

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