Related Experiment Video
Updated: Jul 10, 2026
Generation and Culturing of Primary Human Keratinocytes from Adult Skin
Published on: December 22, 2017
[Research of immunogenity of allogenic keratinocytes]
Jianjun Bi1, Jinjin Wu, Tangyou Zhu
1Department of Dermatology, Research Institute of Field Surgery, Daping Hospital, Third Military Medical University of PLA, Chongqing, 400042, PR China. bjj@mail.tmmu.com.cn
Insights
In vitro cultured keratinocytes show reduced immunogenicity with successive passages. This study demonstrates that passaged human keratinocytes exhibit a gradual decrease in their ability to stimulate allogeneic lymphocytes, suggesting potential for reduced immune rejection in cell-based therapies.
Area of Science:
- Dermatology and immunology research.
- Cell biology and tissue engineering.
Context:
- Understanding keratinocyte immunogenicity is crucial for skin grafting and regenerative medicine.
- In vitro culture conditions can alter the immunological properties of cells.
Purpose:
- To investigate the changes in immunogenicity of human keratinocytes during in vitro culture and serial passaging.
- To quantify the reduction in allogeneic lymphocyte proliferation when co-cultured with passaged keratinocytes.
Summary:
- Human keratinocytes were isolated from foreskins and cultured in vitro up to the fifth passage.
- Immunogenicity was assessed by co-culturing keratinocytes with allogeneic lymphocytes and measuring lymphocyte proliferation.
- Percentages of Langerhans cells and melanocytes decreased significantly with passaging, correlating with reduced lymphocyte proliferation.
Impact:
- The findings suggest that in vitro passaging of keratinocytes leads to a progressive decrease in immunogenicity.
- This reduction in immunogenicity may have implications for improving the success of allogeneic keratinocyte transplantation and reducing immune rejection.
Objective:
To observe the change of the immunogenity of keratinocytes when cultured in vitro.
Methods:
Health children foreskins were digested by dispase and trypase. The human keratinocytes were cultured in vitro and passaged in succession until the fifth passage. Different passage keratinobytes signed by SP method to show the percentage of langerhans cells and melanocytes. Every passage keratinocytes were respectively mixed with allogenic lymphocytes which isolated from peripheral blood, and then the proliferation degree of allogenic lymphocytes was tested.
Results:
Keratinocytes were cultured well in K-SFM medium. When keratinocytes conjugated, every passage cells grew like paving stone. After cryopreservation and then rewarming, the survival exceeded 80%. The percentages of langerhans cells and melanocytes in the primary passage were 5.8% and 8.1% respectively. In the 1st passage they were 2.1% and 2.8% respectively. They were not detected in the second passage. The values of cpm were respectively 482.13 +/- 46.61 (primary passage), 362.50 +/- 35.12 (1st passage), 228.38 +/- 51.46 (2nd passage), 171.86 +/- 34.63 (3rd passage), 143.63 +/- 15.95 (4th passage), and 123.25 +/- 14.39 (5th passage), showing statistically significant differences when compared with control (53.67 +/- 8.61) (P < 0.05). There were statistically significant differences between the primary passage, the 1st passage respectively and the other passages (P < 0.05). There were statistically significant differences between the 4th passage, the 5th passage respectively and the 2nd passage (P < 0.05). There was no statistically significant difference between the 2nd passage and the 3rd passages (P > 0.05). There was not statistically significant difference among the 3rd, the 4th and the 5th passages (P > 0.05).
Conclusion:
Allogenic keratinocytes were cultured in vitro and passaged, and their immunogenity gradually decreased.
Related Concept Videos
01:40Cell-mediated Immune Responses
01:26Antigens Involved in Adaptive Immunity
Complete Antigens
Complete antigens possess both immunogenicity and reactivity.