A comparative study of preparation techniques for improving the viability of striatal grafts using vital stains, in vitro cultures, and in vivo grafts

Cell suspension grafts from embryonic striatal primordia placed into the adult rat striatum survive well and are able to alleviate a number of behavioral deficits caused by excitotoxic lesions to this structure. However, neither the anatomical connectivity between the graft and host nor the functional recovery elicited by the grafts is completely restored. One way in which the survival and function of embryonic striatal grafts may be enhanced is by the improvement of techniques for the preparation of the cell suspension prior to implantation, an issue that has been addressed only to a limited extent. We have evaluated a number of parameters during the preparation procedure, looking at the effects on cell survival over the first 24 h from preparation using vital dyes and the numbers of surviving neurons in vitro, after 4 days in culture, in addition to graft survival and function in vivo. Factors influencing cell survival include the type of trypsinization procedure and the age of donor tissues used for suspension preparation. The presence of DNase has no effect on cell viability but aids the dissociation of the tissue to form single cells. These results have important implications for the use of embryonic striatal grafts in animal models of Huntington's disease, and in any future clinical application of this research.

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A Comparative Study of Preparation Techniques for Improving the Viability of Nigral Grafts using Vital Stains, in Vitro Cultures, and in Vivo Grafts

Cell Transplantation ◽

10.1177/096368979500400204 ◽

1995 ◽

Vol 4 (2) ◽

pp. 173-200 ◽

Cited By ~ 24

Author(s):

Roger A. Barker ◽

Rosemary A. Fricker ◽

D. Nora Abrous ◽

James Fawcett ◽

Stephen B. Dunnett

Keyword(s):

Proteolytic Enzymes ◽

Cell Suspensions ◽

In Vitro Cultures ◽

Preparation Technique ◽

Intracerebral Transplantation ◽

Vital Stains ◽

Long Term Experiment ◽

Dopaminergic Grafts

The intracerebral transplantation of embryonic dopaminergic nigral neurons, although relatively successful, leads to a fairly low yield of surviving cells. Many factors may influence the viability of dopaminergic grafts and one of these is the preparation of the tissue prior to transplantation. We have investigated the effects of different steps during the preparation and storage of embryonic rat nigral cell suspensions on their subsequent survival at a variety of different time points using a combination of techniques and studies. For studies concerned with the first 24 h we employed vital stains, in the period covering the next 7 days we used in vitro cultures, and in the long term experiment we used in vivo grafts. The results suggest that nigral cell suspensions may remain sufficiently viable for grafting for much longer periods than previously reported. In addition a number of parameters which affect cell survival have been characterised, including the age of the embryonic donor tissue, the use of proteolytic enzymes and the trituration procedure used during the preparation of the suspension. The optimal preparation technique, therefore, uses E13-E14 embryos with the dissected ventral mesencephalon being incubated in purified 0.1% trypsin solutions for 60 min and triturated using a flame polished Pasteur pipette. This may have important implications in improving intracerebral transplantation for Parkinson's disease.

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Comparative Study on the Protective Effect of Chlorogenic Acid and 3-(3-Hydroxyphenyl) Propionic Acid against Cadmium-Induced Erythrocyte Cytotoxicity: In Vitro and In Vivo Evaluation

Journal of Agricultural and Food Chemistry ◽

10.1021/acs.jafc.0c04735 ◽

2021 ◽

Vol 69 (13) ◽

pp. 3859-3870

Author(s):

Dai Cheng ◽

Qi Song ◽

Yixin Ding ◽

Qianqian Yu ◽

Yutong Liu ◽

...

Keyword(s):

Comparative Study ◽

Protective Effect ◽

Chlorogenic Acid ◽

Propionic Acid ◽

In Vivo Evaluation

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A comparative study of eicosapentaenoic acid metabolism by human platelets in vivo and in vitro.

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)34360-1 ◽

1985 ◽

Vol 26 (4) ◽

pp. 457-464

Author(s):

C von Schacky ◽

W Siess ◽

S Fischer ◽

P C Weber

Keyword(s):

Comparative Study ◽

Eicosapentaenoic Acid ◽

Acid Metabolism ◽

Human Platelets

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Papaya shoot tip associated endophytic bacteria isolated from in vitro cultures and host–endophyte interaction in vitro and in vivo

Canadian Journal of Microbiology ◽

10.1139/w06-141 ◽

2007 ◽

Vol 53 (3) ◽

pp. 380-390 ◽

Cited By ~ 36

Author(s):

Pious Thomas ◽

Sima Kumari ◽

Ganiga K. Swarna ◽

T.K.S. Gowda

Keyword(s):

Culture Medium ◽

Carica Papaya ◽

In Vitro Cultures ◽

Dependent Manner ◽

16S Rdna Sequence ◽

16S Rdna Sequence Analysis ◽

Single Organism ◽

Dose Dependent

Fourteen distinct bacterial clones were isolated from surface-sterilized shoot tips (~1 cm) of papaya (Carica papaya L. ‘Surya’) planted on Murashige and Skoog (MS)-based papaya culture medium (23/50 nos.) during the 2–4 week period following in vitro culturing. These isolates were ascribed to six Gram-negative genera, namely Pantoea ( P. ananatis ), Enterobacter ( E. cloacae ), Brevundimonas ( B. aurantiaca ), Sphingomonas , Methylobacterium ( M. rhodesianum ), and Agrobacterium ( A. tumefaciens ) or two Gram-positive genera, Microbacterium ( M. esteraromaticum ) and Bacillus ( B. benzoevorans ) based on 16S rDNA sequence analysis. Pantoea ananatis was the most frequently isolated organism (70% of the cultures) followed by B. benzoevorans (13%), while others were isolated from single stocks. Bacteria-harboring in vitro cultures often showed a single organism. Pantoea, Enterobacter, and Agrobacterium spp. grew actively on MS-based normal papaya medium, while Microbacterium, Brevundimonas, Bacillus, Sphingomonas, and Methylobacterium spp. failed to grow in the absence of host tissue. Supplying MS medium with tissue extract enhanced the growth of all the organisms in a dose-dependent manner, indicating reliance of the endophyte on its host. Inoculation of papaya seeds with the endophytes (20 h at OD550 = 0.5) led to delayed germination or slow seedling growth initially. However, the inhibition was overcome by 3 months and the seedlings inoculated with Pantoea, Microbacterium, or Sphingomonas spp. displayed significantly better root and shoot growths.

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