Development of a chemically defined serum- and protein-free medium for growth of human peripheral lymphocytes.

Shive, William; Pinkerton, Frederick D.; Humphreys, John F.; Johnson, Margaret M.; Hamilton, W G; Matthews, Kathleen S.

doi:10.1073/pnas.83.1.9

Cited by 32 publications

(8 citation statements)

References 20 publications

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“…The functional assessment of intracellular deficiencies of micronutrients was performed by the lymphocyte transformation assay developed by Shive et al(19–24). In this procedure, blood is collected in two 10ml cell preparation tubes (Becton Dickinson) containing sodium citrate and ficoll hypaque.…”

Section: Methodsmentioning

confidence: 99%

Coenzyme Q10 deficiency in patients with Parkinson's disease

Mischley

Allen

Bradley

2012

Journal of the Neurological Sciences

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Reactive oxygen species (ROS) are well known to contribute to the pathophysiology of Parkinson’s disease (PD). Clinical trials of antioxidants are currently underway in PD patients, however, antioxidant research has been hindered by a lack of peripheral biomarkers. METHODS Twenty-two patients with PD elected to have a novel antioxidant assessment (Functional Intracellular Assay (FIA), Spectra Cell Lab, Houston, TX) performed between 2004–2008. Each PD case was compared to four age- and gender- matched controls (n=88) in four separate, random iterations using laboratory data submitted during the same time period. Logistic regression was used to determine the odds of functional deficiency in antioxidant nutrients (i.e., glutathione, coenzyme Q10, selenium, vitamin E and alpha-lpoic acid) by case-control status. The proportion of cases with functional deficiency was also compared to that for controls by chi2 test. RESULTS Compared to cases, PD patients had a significantly greater odds of deficiency in coenzyme Q10 status (OR: 4.7–5.4; 95% CI: 1.5–17.7; P = 0.003–0.009) based on FIA results, but not of vitamin E, selenium, lipoic acid, or glutathione (all P>0.05). The proportion of cases with coenzyme Q10 deficiency was also significantly greater in cases than in controls (32–36% vs. 8–9%; P=0.0012–0.006). CONCLUSIONS Deficiency of coenzyme Q10 assessed via FIA should be explored as a potential peripheral biomarker of antioxidant status in PD.

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Section: Methodsmentioning

confidence: 99%

Coenzyme Q10 deficiency in patients with Parkinson's disease

Mischley

Allen

Bradley

2012

Journal of the Neurological Sciences

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“…Earlier, we made an observation that IMDM is inferior to a-medium for serum-free culture of human marrow progenitors plated at low cell concentrations. Recently, Shive et al [17] described a chemically defined, serum-and protein-free medium (CFBI 1OOO) for suspension culture of human peripheral lymphocytes. The concentrations of individual components of CFBI 1 ,OOO reflect minimal effective concentrations and are significantly lower than those of IMDM and a-medium.…”

Section: Introductionmentioning

confidence: 99%

Serum‐free culture of human hemopoietic progenitors in attenuated culture media

Sonoda

Ogawa

1988

American J Hematol

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To elucidate the precise mechanisms of molecular and cellular regulation of hemopoiesis, it is necessary to develop a chemically defined culture assay for purified hemopoietic progenitors. To approach this long-term goal, we attempted to develop a serum-free culture system for enriched human progenitors that permits expression of all hemopoietic lineages and stages of development. Preliminary studies indicated that alpha-medium was superior to Iscove's modified Dulbecco's medium (IMDM) and that culture under low (5%) oxygen condition was better than an ambient level of oxygen. We developed an attenuated (modified quarter-strength) alpha-medium and compared the colony-supporting ability of the three media by plating 1,000 bone marrow null cells per dish in the presence of a combination of recombinant human colony-stimulating factors (CSFs). The numbers of colonies supported in alpha-medium and attenuated alpha-medium were approximately 70% of those in serum-containing cultures. IMDM failed to support colony formation. While, in general, the colony sizes were smaller in the serum-free cultures than in the serum-containing cultures, a variety of types of single lineage and multilineage colonies were seen in serum-free culture. A linear relationship between cell number and colony formation was seen in 100-2,000 cells per dish. Serum-free cultures of enriched human progenitors should be an important tool for analysis of the mechanisms of recombinant CSFs.

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“…Lymphocyte growth in culture media to which individual micronutrients are added or deleted and compared to the lymphocyte growth in optimal culture media are determined. In the study reported here, the influence of the EMF is the difference between samples taken before and after exposure to the EMF (Tu et al, 1994;Shrive et al, 1986;Boerner, 2001;Shive, 1988). Thus, each individual serves as his/her own control as the lymphocytes were cultured from the patients blood taken before and after exposure to the EMF of the THERAMAG device.…”

Section: Spectracell Studiesmentioning

confidence: 99%

Reduction of Pain in Reflex Sympathetic Dystrophy (RSD) Associated with Objective Biochemical Changes in Circulating Lymphocytes

Ericsson

Hazlewood²,

Markov

et al. 2005

Environmentalist

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In an effort to explain the benefit of therapeutic use of electromagnetic fields (EMF) a systemic effect has been proposed by us. To assess the efficacy of this approach, an objective biochemical approach was developed. Ten patients with Reflex Sympathetic Dystrophy (RSD) were clinically evaluated, and lymphocytes were isolated from their blood samples obtained before and after exposure of the uninvolved limb to an EMF. Utilizing a SpectraCell method that includes radio labeled molecules and protein-free media for culturing the lymphocytes, an elevation of the content of fructose, serine, glycine, and calcium cellular metabolic uptake were found following in the culture to EMF in comparison with non-exposed lymphocytes ( p < 0.01). In addition, the pain level was determined by a conventional visual analogue scale (VAS) before and after EMF exposure, evidencing a significant pain relief. Specifically, after exposure to 120 pps semi sinewave, 1500Gauss EMF, generated by a THERAMAG device, an improvement in the flexibility of the limb and a reduction in swelling of the affected extremity were detected clinically. These findings are in concert with the new hypothesis that, with relief of pain, lymphocytes are predominately altered in their cell cycle from M phase to S phases associated with increased structuring of intracellular water. A consideration of the basic understanding of the role lymphocytes may be inferred from this preliminary study. An extensive review of the literature on the basic science and therapeutic use of magnetic fields in humans is provided in an attempt to understand the relevance of magnetic therapy of specific pain syndromes.

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Development of a chemically defined serum- and protein-free medium for growth of human peripheral lymphocytes.

Cited by 32 publications

References 20 publications

Coenzyme Q10 deficiency in patients with Parkinson's disease

Coenzyme Q10 deficiency in patients with Parkinson's disease

Serum‐free culture of human hemopoietic progenitors in attenuated culture media

Reduction of Pain in Reflex Sympathetic Dystrophy (RSD) Associated with Objective Biochemical Changes in Circulating Lymphocytes

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