Preliminary investigation of human lymphatic malformations in vitro

Yuemeng Dai, Fang Hou, Ali Saad, Chun Yang Fan, Lisa M. Buckmiller, James Y. Suen, Gresham T. Richter

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Purpose: To develop an in vitro model of human lymphatic malformations (LM) that reflects histological characteristics of native LM. Methods: Fresh human LM (n = 6) were harvested, sectioned, explanted into a fibrinogen gel, and cultured. A total of 25 explants were developed and observed for primary and peripheral cellular growth. On days 9 to 10, the cultured tissues and gels were collected and fixed in 10% formalin. Primary LM and surrounding gel matrix were sectioned and stained for H&E analysis. Immunohistochemistry was performed for Prox-1 and D2-40, known markers for lymphatic endothelium, and Ki-67, a marker of cellular proliferation. Results: On culture day 3, cells were observed to grow into the gel surrounding the primary tissue explants. Persistent and significant growth into the gel matrix was observed for each specimen at subsequent measurement intervals (day 6 and day 10, P <.0001). H&E staining of all the LM explants demonstrated survival and intact organization and cellular structure reflective of the original LM specimens. Microchannels were observed in the surrounding gel suggesting the presence of newly formed lymphatic vessels. Positive-immunohistochemical staining for D2-40 and Prox-1 revealed organized lymphatic endothelia within each specimen and associated microchannels distal to the explants in the gel matrix. Scattered cells in the gel matrix stained positive for Ki-67. Conclusions: This experimental model suggests that human LM can be preserved and observed to grow in vitro with structural characteristics, and immunohistologic qualities similar to native LM. This model may provide a facile tool for basic and translational research on LM.

Original languageEnglish (US)
Pages (from-to)2435-2442
Number of pages8
JournalLaryngoscope
Volume121
Issue number11
DOIs
StatePublished - Nov 1 2011
Externally publishedYes

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Gels
Lymphatic Endothelium
Naproxen
Staining and Labeling
Lymphatic Vessels
Translational Medical Research
In Vitro Techniques
Cellular Structures
Growth
Fibrinogen
Formaldehyde
Theoretical Models
Immunohistochemistry
Cell Proliferation
Survival

All Science Journal Classification (ASJC) codes

  • Otorhinolaryngology

Cite this

Dai, Y., Hou, F., Saad, A., Fan, C. Y., Buckmiller, L. M., Suen, J. Y., & Richter, G. T. (2011). Preliminary investigation of human lymphatic malformations in vitro. Laryngoscope, 121(11), 2435-2442. https://doi.org/10.1002/lary.22187

Preliminary investigation of human lymphatic malformations in vitro. / Dai, Yuemeng; Hou, Fang; Saad, Ali; Fan, Chun Yang; Buckmiller, Lisa M.; Suen, James Y.; Richter, Gresham T.

In: Laryngoscope, Vol. 121, No. 11, 01.11.2011, p. 2435-2442.

Research output: Contribution to journalArticle

Dai, Y, Hou, F, Saad, A, Fan, CY, Buckmiller, LM, Suen, JY & Richter, GT 2011, 'Preliminary investigation of human lymphatic malformations in vitro', Laryngoscope, vol. 121, no. 11, pp. 2435-2442. https://doi.org/10.1002/lary.22187
Dai Y, Hou F, Saad A, Fan CY, Buckmiller LM, Suen JY et al. Preliminary investigation of human lymphatic malformations in vitro. Laryngoscope. 2011 Nov 1;121(11):2435-2442. https://doi.org/10.1002/lary.22187
Dai, Yuemeng ; Hou, Fang ; Saad, Ali ; Fan, Chun Yang ; Buckmiller, Lisa M. ; Suen, James Y. ; Richter, Gresham T. / Preliminary investigation of human lymphatic malformations in vitro. In: Laryngoscope. 2011 ; Vol. 121, No. 11. pp. 2435-2442.
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abstract = "Purpose: To develop an in vitro model of human lymphatic malformations (LM) that reflects histological characteristics of native LM. Methods: Fresh human LM (n = 6) were harvested, sectioned, explanted into a fibrinogen gel, and cultured. A total of 25 explants were developed and observed for primary and peripheral cellular growth. On days 9 to 10, the cultured tissues and gels were collected and fixed in 10{\%} formalin. Primary LM and surrounding gel matrix were sectioned and stained for H&E analysis. Immunohistochemistry was performed for Prox-1 and D2-40, known markers for lymphatic endothelium, and Ki-67, a marker of cellular proliferation. Results: On culture day 3, cells were observed to grow into the gel surrounding the primary tissue explants. Persistent and significant growth into the gel matrix was observed for each specimen at subsequent measurement intervals (day 6 and day 10, P <.0001). H&E staining of all the LM explants demonstrated survival and intact organization and cellular structure reflective of the original LM specimens. Microchannels were observed in the surrounding gel suggesting the presence of newly formed lymphatic vessels. Positive-immunohistochemical staining for D2-40 and Prox-1 revealed organized lymphatic endothelia within each specimen and associated microchannels distal to the explants in the gel matrix. Scattered cells in the gel matrix stained positive for Ki-67. Conclusions: This experimental model suggests that human LM can be preserved and observed to grow in vitro with structural characteristics, and immunohistologic qualities similar to native LM. This model may provide a facile tool for basic and translational research on LM.",
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