| Autor: |
N. Sharma, M. Madhan Kumar, K. K. Mohanty, V. Khandelwal |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Re:GEN Open, Vol 2, Iss 1, Pp 145-149 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2766-2705 |
| DOI: |
10.1089/REGEN.2022.0014 |
| Popis: |
Background: For studying in vitro immune responses, peripheral blood mononuclear cells (PBMCs) and its immune cell components isolated from human blood are used. In this study, we have standardized the appropriate percentage of human AB serum (HABS) for a cost-effective work layout in monocyte cultures derived from human PBMC. Methodology: For studying the usage of AB serum in culture, PBMCs-derived monocyte-derived macrophages (MDMs) were studied in healthy individuals (purified protein derivative (PPD) positive (n?=?3), PPD negative (n?=?3)) and pulmonary tuberculosis patients (PTB) (n?=?3). Monocytes derived by density gradient centrifugation were adhered and differentiated into MDMs for the study. The morphology and viability of the MDMs in different percentages of AB sera (5% and 10% HABS) were assessed for optimization. Results: No significant difference was observed in the usage of 5% and 10% of HABS in MDMs culture in terms of morphology and viability in different groups of study subjects: PPD positive subjects (P value: 1.0000), PPD negative subjects (P value: 0.6531), and PTB (P value: 0.8195)), and 5% HABS was found to be optimum for culturing MDMs. Conclusion: This work is novel because to our knowledge, none of the studies have compared different percentages of human sera for evaluating their efficiency in cell culture. Since the cost of HABS is about four to five times more than that of fetal bovine serum, a study of this kind that explores the optimum usage of serum for supporting cell growth without compromising cell viability and morphology will be of much use in cell culture studies. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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