A Mixed Lymphocyte Culture Micro‐Technique and its Application in Different Family and Unrelated Combinations

Bondevik, H; Helgesen, Astri; Thoresen, A. B.; Thorsby, E.

doi:10.1111/j.1399-0039.1974.tb00279.x

Cited by 20 publications

(4 citation statements)

References 7 publications

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“…On several occasions, cultures were harvested on both days 5 and 6. The latter gave somewhat higher stimulation, similar to that seen when lymphocytes are used (Bondevik et al 1974). Stimulation kinetics for MLC and MLMC were similar through days 1 to 6.…”

Section: Dose Response and Kinetics Of Macrophage Stimulationsupporting

confidence: 70%

“…If not otherwise stated, 12,500 stimulating macrophages were used, and cultures were set up as triplicates and harvested by a semi-automatic harvesting machine (Skatron) on day 5, after an 18 h pulse of H-thymidine. The variation among the triplicates, expressed as s.e., was usually less than 20% of the mean, but was often greater than in our MLC cultures (Bondevik et al 1974). Therefore, we were sometimes compelled to disregard the result of a single culture if it was disparate from the two others of a triplicate (obviously due to technical failure).…”

Section: Mixed Lymphocyte Macrophage Cultures (Mlmc)mentioning

confidence: 87%

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Stimulation of Human Lymphocytes by Allogenic Macrophages in vitro

Kaakinen¹,

Hirschberg

1977

Tissue Antigens

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Macrophages were obtained from human peripheral blood by incubating mononuclear cells in plastic tissue culture flasks. After 1—14 days, the cells were used in mixed lymphocyte macrophage cultures. Macrophages could not, themselves, be stimulated to proliferation by allogenic cells, but stimulated allogenic lymphocytes. By the use of responding cells and stimulating macrophages from HLA‐D homozygous individuals, the HLA‐D determinants could be shown to be responsible for the stimulation of allogenic lymphocytes.

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Section: Dose Response and Kinetics Of Macrophage Stimulationsupporting

confidence: 70%

Section: Mixed Lymphocyte Macrophage Cultures (Mlmc)mentioning

confidence: 87%

Stimulation of Human Lymphocytes by Allogenic Macrophages in vitro

Kaakinen¹,

Hirschberg

1977

Tissue Antigens

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“…The problems associated with the expression of the results of MLC experiments were thoroughly discussed in the Oslo Workshop (see Thorsby et al 1974).…”

Section: Discussionmentioning

confidence: 99%

Comparison of Microtitre Plates With Flat‐bottomed and Round‐bottomed Wells for Mixed Lymphocyte Culture (Mlc)

Herva¹

1977

Acta Pathologica Microbiologica Scandinavica Section C Immunolo

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To compare microtitre plates with flat‐bottomed and round‐bottomed wells and to standardize a method for mixed lymphocyte culture (MLC), the effects of cell number, culture time, ?3H‐thymidine concentration and labelling time were studied. On both plates, allogeneic cells induced increased RNA synthesis beginning at about 30 hours and increased DNA synthesis beginning at about 50 hours, if suitable cell numbers were used. On plates with flat‐bottomed wells, 1.5 × 105 responding and stimulating cells per well had near‐exponential growth on day four and five, often through day six; on plates with round‐bottomed wells the corresponding cell number was 0.25–1.0 (optimally 0.5) × 105. Near these cell numbers, the response depended closely on the number of responding cells. On plates with flat‐bottomed wells, stimulating cells had a dose‐dependent effect on the response, whereas on plates with round‐bottomed wells, increasing the stimulating cell dose did not consistently strengthen the response. Both plate types proved suitable for MLC experiments; plates with round‐bottomed wells have the obvious advantage of requiring smaller cell numbers. 3H‐thymidine (spec, act 2000 mCi/mmol) self‐suppressed its incorporation, which increased only slightly or even decreased if labelling time exceeded 12–18 hours. Relative responses remained virtually unaltered, however, with 3H‐concentrations of 0.5 and 2.0 μCi/ml and with labelling times of 8 and 24 hours.

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“…n.t. not tested Table 2 The typing cells used in this study MM PF EA IB MS Kaakinen et al 1975Bondevik et al 1974 This article OH PM K T 2,12/2,12 2,12/2,12 2,W5/3,W5…”

Section: T H E Typing Cellsmentioning

confidence: 99%

Identification of Five Lymphocyte Activating Determinants in Man

et al. 1975

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Eleven putative LD‐1 locus homozygous cell donors were found by MLC tests in non‐con‐sanguineous families. Lymphocytes from each of them were used as stimulating typing cells in MLC! tests together with responding cells from the other homozygous cell donors and from 45 random unrelated individuals. Four of the stimulating typing cells were found to identify a determinant in non‐random association with IIL‐.A7; LD‐ 7a, three other cells typed for a determinant non‐randomly associated with HL‐A8; LD‐8a, and one cell seems to identify LD‐W5 in non‐random association with W5. Two possible additional LD‐1 determinants, LD‐oh and LD‐pm, were less well defined with two of the cells. The combined gene frequency of the five possible LD‐1 determinants is approximately 0.45 among Norwegians.

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A Mixed Lymphocyte Culture Micro‐Technique and its Application in Different Family and Unrelated Combinations

Cited by 20 publications

References 7 publications

Stimulation of Human Lymphocytes by Allogenic Macrophages in vitro

Stimulation of Human Lymphocytes by Allogenic Macrophages in vitro

Comparison of Microtitre Plates With Flat‐bottomed and Round‐bottomed Wells for Mixed Lymphocyte Culture (Mlc)

Identification of Five Lymphocyte Activating Determinants in Man

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