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Effect of LAK cells against three-dimensional tumor tissue. In vitro study using multi-cellular human glioma
J Jääskeläinen1, P Kalliomäki, A Paetau
1Department of Neurosurgery, Helsinki University Central Hospital, Finland.
Abstract:
The anti-tumor mechanisms of local LAK cell therapy are difficult to study in vivo. We describe a method to study in vitro the action of LAK cells against three-dimensional tumor tissue. Spherical cell aggregates (spheroids) grown from human glioma cell lines H-2 and U-251 were labeled with 51Cr and then incubated for up to 24 h with LAK cells. After the incubation, most spheroids were still macroscopically identifiable, and the measured reduction of volume did not correlate to the extent of damage. LAK cells infiltrated into spheroid tissue slowly as a frontier which explains why the specific 51Cr release was clearly slower from spheroids than corresponding single cell suspensions. The infiltrated area was at 1 to 2 h very thin but by 8 to 12 h consisted already of several cell layers. Most H-2 spheroids became totally infiltrated by 16 to 24 h whereas in U-251 spheroids the infiltration usually remained peripheral. In accordance with the different extent of infiltration, H-2 spheroids were clearly more sensitive to LAK cells than U-251 spheroids: at E/T ratio 10:1 the mean specific 51Cr release by 24 h was 63 and 36%, respectively. A single exposure to LAK cells released 51Cr from H-2 spheroids approximately 12 h but over 24 h from U-251 spheroids. The spheroid model can be used to study the infiltrative capacity and cytotoxicity of LAK cells against three-dimensional tumor tissue, and the method may help to find an optimal mode of local LAK cell therapy, i.e., proper combination of lymphokines and LAK cells, and proper timing of their administration.
Insights
This study introduces a novel in vitro method using 3D tumor spheroids to assess LAK cell therapy effectiveness. The spheroid model accurately reflects LAK cell infiltration and cytotoxicity against solid tumors.
Area of Science:
- Oncology
- Immunotherapy
- Cell Biology
Background:
- Studying local LAK (Lymphokine-Activated Killer) cell therapy's anti-tumor effects in vivo is challenging.
- A new in vitro model is needed to investigate LAK cell interactions with three-dimensional (3D) tumor tissue.
Purpose of the Study:
- To develop and validate an in vitro method for evaluating LAK cell action against 3D tumor spheroids.
- To assess LAK cell infiltrative capacity and cytotoxicity in a 3D tumor microenvironment.
Main Methods:
- Human glioma cell lines (H-2 and U-251) were cultured as 3D spheroids.
- Spheroids were labeled with 51Cr and incubated with LAK cells for up to 24 hours.
- LAK cell infiltration and 51Cr release were quantified to measure cytotoxicity.
Main Results:
- LAK cell infiltration into spheroids was slow and frontier-like, differing from single-cell suspensions.
- H-2 spheroids showed greater sensitivity to LAK cells (63% 51Cr release at 24h) than U-251 spheroids (36% 51Cr release).
- Infiltration depth and speed varied between H-2 and U-251 spheroids, impacting LAK cell efficacy.
Conclusions:
- The 3D spheroid model effectively simulates LAK cell interaction with solid tumor tissue.
- This model aids in optimizing local LAK cell therapy by determining optimal lymphokine combinations and administration timing.
- The findings provide insights into LAK cell therapy for solid tumors, particularly gliomas.