Journal of Saveh University of Medical Sciences

Enhancing Karyotyping Success: The Role of Conditioning Media in Modulating Cell Proliferation and Apoptosis in Lymphopenic Samples

Document Type : Original Article

Authors

Bentolhoda Kuhestani dehaghi 1
maryam panahi 2
Amir abbas Navidinia 1
Hamideh Rahmani seraji 3
Elham Jafaei 4
mozhgan amirpour 1
Mehdi Allahbakhshian farsani 1

1 Department of Hematology and Blood Banking, Faculty of allied medical sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.

3 Department of Hematology and Oncology, Taleghani Hospital, Shahid Beheshti University of Medical Science, Tehran, Iran.

4 Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

https://doi.org/10.22034/jsavehums.2025.225628
Abstract
Introduction: Conventional karyotyping (CK) fundamentally relies on a sufficient quantity of viable and highly proliferative cells, a requirement that frequently leads to culture failure—particularly under conditions of reduced cellularity due to physiological or pathological lymphopenia. In such cases, inadequate cell interactions and suboptimal proliferation within the culture medium compromise diagnostic yield. This study investigated the impact of Conditioning Media (CM) on cell survival, proliferation, and apoptosis in lymphopenic samples.
Methods: To closely emulate in vivo microenvironmental conditions, CM was employed to optimize the in vitro culture system. Data were obtained through mitotic index (MI) assessments, proliferation assays, and apoptosis analyses, supplemented by RT-qPCR to examine dynamic changes in gene expression associated with cell cycle regulation and survival pathways.
Results: The incorporation of CM into the culture system significantly improved culture success rates. This was demonstrated by enhanced cell survival, increased mitotic indices, elevated Ki-67 expression—indicative of heightened proliferative activity—and a pronounced progression into the S phase of the cell cycle. Concurrently, a marked reduction in apoptosis was observed, suggesting improved cellular viability and resilience.
Conclusion: Our findings support the integration of CM into standard culture protocols to improve growth conditions, particularly in specimens with low cell counts or limited proliferative capacity—factors that traditionally impede successful karyotypic analysis. These improvements highlight the transformative potential of CM in refining CK methodologies, thereby broadening their reliability and applicability in clinical genetic diagnostics.

Keywords

conventional karyotype
conditioning media
Lymphopenia
Journal of Saveh University of Medical Sciences
Volume 1, Issue 1
Spring 2025
Pages 12-25

Home

Guide for Authors

Submit Manuscript

Reviewers

Contact Us