Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice

Dorte Bratbo Sørensen; Stine Broeng Metzdorff; Louise Kruse Jensen; Kamilla H Andersen; Anne Charlotte Teilmann; Henrik Elvang Jensen; Hanne Frøkiær

doi:10.30802/AALAS-JAALAS-18-000064

Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice

Dorte Bratbo Sørensen, Stine Broeng Metzdorff, Louise Kruse Jensen, Kamilla H Andersen, Anne Charlotte Teilmann, Henrik Elvang Jensen, Hanne Frøkiær

Section of Research and Education

3 Citations (Scopus)

Abstract

We compared 6 frequently used mouse blood-sampling methods (lateral tail incision; tail-tip amputation; sublingual, submandibular, and saphenous vein puncture; and retrobulbar sinus puncture during isoflurane anesthesia) with regard to induction of local and systemic inflammation, stomach contents, weight changes, and corticosterone levels at 6 h to 12 d after sampling. Local inflammation was assessed through histopathology and assessment of the expression of inflammation and tissue damage–related genes (S1008/9A, Cxcl2, Il1b, Nlrp3, Il6, and Il33) in sampled tissue. Systemic inflammation was assessed through quantification of plasma haptoglobin levels, measurement of blood Il1b expression, and evaluation of histopathologic changes in lung, kidney, liver, and spleen. Apart from slight, transient increases in plasma haptoglobin levels after lateral tail incision, retrobulbar sinus puncture, and saphenous vein puncture, no other signs of systemic inflammation were found. Mice subjected to retrobulbar sinus puncture, sublingual puncture, or isoflurane anesthesia only showed the highest plasma corticosterone concentrations. Retrobulbar sinus puncture had the largest effect on body weight loss. Retrobulbar sinus puncture, sublingual puncture, and submandibular puncture only showed minor and in, most cases, fastresolving inflammation. By contrast, blood sampling by lateral tail incision, tail-tip amputation, or saphenous vein puncture caused tissue damage and inflammation locally at the sampling site, which resolved more slowly compared with head-region sampling techniques, according to results from pathologic and gene expression assessments. Expression of S1008/9A, Cxcl2, Il1b, and Nlrp3 increased 10- to 1000-fold and did not return to baseline until day 6 after sampling or later and did not resolve after tail-tip amputation within the 12-d observation period. Increased expression of genes involved in inflammation and tissue repair correlated with histopathologic changes and may thus represent a quantitative supplement to histopathology. In conclusion, none of the tested methods for obtaining blood samples from mice is superior, according to simultaneous immunologic, histopathologic, and animal welfare–related parameters.

Original language	English
Journal	JAALAS - Journal of the American Association for Laboratory Animal Science
Volume	58
Issue number	3
Pages (from-to)	362-372
ISSN	1559-6109
DOIs	https://doi.org/10.30802/AALAS-JAALAS-18-000064
Publication status	Published - May 2019

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Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice. / Sørensen, Dorte Bratbo; Metzdorff, Stine Broeng; Jensen, Louise Kruse et al.
In: JAALAS - Journal of the American Association for Laboratory Animal Science, Vol. 58, No. 3, 05.2019, p. 362-372.

Research output: Contribution to journal › Journal article › Research › peer-review

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title = "Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice",

abstract = "We compared 6 frequently used mouse blood-sampling methods (lateral tail incision; tail-tip amputation; sublingual, submandibular, and saphenous vein puncture; and retrobulbar sinus puncture during isoflurane anesthesia) with regard to induction of local and systemic inflammation, stomach contents, weight changes, and corticosterone levels at 6 h to 12 d after sampling. Local inflammation was assessed through histopathology and assessment of the expression of inflammation and tissue damage–related genes (S1008/9A, Cxcl2, Il1b, Nlrp3, Il6, and Il33) in sampled tissue. Systemic inflammation was assessed through quantification of plasma haptoglobin levels, measurement of blood Il1b expression, and evaluation of histopathologic changes in lung, kidney, liver, and spleen. Apart from slight, transient increases in plasma haptoglobin levels after lateral tail incision, retrobulbar sinus puncture, and saphenous vein puncture, no other signs of systemic inflammation were found. Mice subjected to retrobulbar sinus puncture, sublingual puncture, or isoflurane anesthesia only showed the highest plasma corticosterone concentrations. Retrobulbar sinus puncture had the largest effect on body weight loss. Retrobulbar sinus puncture, sublingual puncture, and submandibular puncture only showed minor and in, most cases, fastresolving inflammation. By contrast, blood sampling by lateral tail incision, tail-tip amputation, or saphenous vein puncture caused tissue damage and inflammation locally at the sampling site, which resolved more slowly compared with head-region sampling techniques, according to results from pathologic and gene expression assessments. Expression of S1008/9A, Cxcl2, Il1b, and Nlrp3 increased 10- to 1000-fold and did not return to baseline until day 6 after sampling or later and did not resolve after tail-tip amputation within the 12-d observation period. Increased expression of genes involved in inflammation and tissue repair correlated with histopathologic changes and may thus represent a quantitative supplement to histopathology. In conclusion, none of the tested methods for obtaining blood samples from mice is superior, according to simultaneous immunologic, histopathologic, and animal welfare–related parameters.",

author = "S{\o}rensen, {Dorte Bratbo} and Metzdorff, {Stine Broeng} and Jensen, {Louise Kruse} and Andersen, {Kamilla H} and Teilmann, {Anne Charlotte} and Jensen, {Henrik Elvang} and Hanne Fr{\o}ki{\ae}r",

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doi = "10.30802/AALAS-JAALAS-18-000064",

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T1 - Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice

AU - Sørensen, Dorte Bratbo

AU - Metzdorff, Stine Broeng

AU - Jensen, Louise Kruse

AU - Andersen, Kamilla H

AU - Teilmann, Anne Charlotte

AU - Jensen, Henrik Elvang

AU - Frøkiær, Hanne

PY - 2019/5

Y1 - 2019/5

N2 - We compared 6 frequently used mouse blood-sampling methods (lateral tail incision; tail-tip amputation; sublingual, submandibular, and saphenous vein puncture; and retrobulbar sinus puncture during isoflurane anesthesia) with regard to induction of local and systemic inflammation, stomach contents, weight changes, and corticosterone levels at 6 h to 12 d after sampling. Local inflammation was assessed through histopathology and assessment of the expression of inflammation and tissue damage–related genes (S1008/9A, Cxcl2, Il1b, Nlrp3, Il6, and Il33) in sampled tissue. Systemic inflammation was assessed through quantification of plasma haptoglobin levels, measurement of blood Il1b expression, and evaluation of histopathologic changes in lung, kidney, liver, and spleen. Apart from slight, transient increases in plasma haptoglobin levels after lateral tail incision, retrobulbar sinus puncture, and saphenous vein puncture, no other signs of systemic inflammation were found. Mice subjected to retrobulbar sinus puncture, sublingual puncture, or isoflurane anesthesia only showed the highest plasma corticosterone concentrations. Retrobulbar sinus puncture had the largest effect on body weight loss. Retrobulbar sinus puncture, sublingual puncture, and submandibular puncture only showed minor and in, most cases, fastresolving inflammation. By contrast, blood sampling by lateral tail incision, tail-tip amputation, or saphenous vein puncture caused tissue damage and inflammation locally at the sampling site, which resolved more slowly compared with head-region sampling techniques, according to results from pathologic and gene expression assessments. Expression of S1008/9A, Cxcl2, Il1b, and Nlrp3 increased 10- to 1000-fold and did not return to baseline until day 6 after sampling or later and did not resolve after tail-tip amputation within the 12-d observation period. Increased expression of genes involved in inflammation and tissue repair correlated with histopathologic changes and may thus represent a quantitative supplement to histopathology. In conclusion, none of the tested methods for obtaining blood samples from mice is superior, according to simultaneous immunologic, histopathologic, and animal welfare–related parameters.

AB - We compared 6 frequently used mouse blood-sampling methods (lateral tail incision; tail-tip amputation; sublingual, submandibular, and saphenous vein puncture; and retrobulbar sinus puncture during isoflurane anesthesia) with regard to induction of local and systemic inflammation, stomach contents, weight changes, and corticosterone levels at 6 h to 12 d after sampling. Local inflammation was assessed through histopathology and assessment of the expression of inflammation and tissue damage–related genes (S1008/9A, Cxcl2, Il1b, Nlrp3, Il6, and Il33) in sampled tissue. Systemic inflammation was assessed through quantification of plasma haptoglobin levels, measurement of blood Il1b expression, and evaluation of histopathologic changes in lung, kidney, liver, and spleen. Apart from slight, transient increases in plasma haptoglobin levels after lateral tail incision, retrobulbar sinus puncture, and saphenous vein puncture, no other signs of systemic inflammation were found. Mice subjected to retrobulbar sinus puncture, sublingual puncture, or isoflurane anesthesia only showed the highest plasma corticosterone concentrations. Retrobulbar sinus puncture had the largest effect on body weight loss. Retrobulbar sinus puncture, sublingual puncture, and submandibular puncture only showed minor and in, most cases, fastresolving inflammation. By contrast, blood sampling by lateral tail incision, tail-tip amputation, or saphenous vein puncture caused tissue damage and inflammation locally at the sampling site, which resolved more slowly compared with head-region sampling techniques, according to results from pathologic and gene expression assessments. Expression of S1008/9A, Cxcl2, Il1b, and Nlrp3 increased 10- to 1000-fold and did not return to baseline until day 6 after sampling or later and did not resolve after tail-tip amputation within the 12-d observation period. Increased expression of genes involved in inflammation and tissue repair correlated with histopathologic changes and may thus represent a quantitative supplement to histopathology. In conclusion, none of the tested methods for obtaining blood samples from mice is superior, according to simultaneous immunologic, histopathologic, and animal welfare–related parameters.

U2 - 10.30802/AALAS-JAALAS-18-000064

DO - 10.30802/AALAS-JAALAS-18-000064

M3 - Journal article

C2 - 30947767

SN - 1559-6109

VL - 58

SP - 362

EP - 372

JO - JAALAS - Journal of the American Association for Laboratory Animal Science

JF - JAALAS - Journal of the American Association for Laboratory Animal Science

IS - 3

ER -

Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice

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