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All Journal Medical Journal of Indonesia

Caroline T. Sardjono

Faculty of Medicine, Maranatha Christian University, Bandung

Author-ID : 372299

Medicine & Pharmacology

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Application of a modified method for stem cell isolation from lipoaspirates in a basic lab Sardjono, Caroline T.; Setiawan, Melina; Frisca, Frisca; Saputra, Virgi; Aniko, Gwendy; Sandra, Ferry
Medical Journal of Indonesia Vol 18, No 2 (2009): April-June
Publisher : Faculty of Medicine Universitas Indonesia

Aim Lipoaspirate, a wasted by product from liposuction procedure recently has been shown to contain abundant mesenchymal stem cells (MSCs). MSCs have been studied in many research areas to regenerate many cell lineages including, myogenic, cardiomyogenic, and angiogenic lineages. The large quantity of MSCs in lipoaspirate, makes it an attractive source for stem cells used in research and clinical applications. A simplified method which is suitable to be performed in a basic laboratory will facilitate development of stem cell research in developing countries. Therefore the outcomes from this study are expected to encourage the progress of stem cell research in Indonesia.Methods Lipoaspirate was digested using collagenase type I, followed by a basic filtration method. Purification of MSCs was done by cell culture for 2-3 days followed by supernatant removal. To confirm the homogenous population of MSCs, an analysis using flowcytometry was performed based on the MSCs minimal criteria developed by Mesenchymal and Tissue Stem Cell Committee of the International Society of Cell Therapy.Resuts MSCs were able to be obtained at 16.41 Â± 8.22 x 108 cells per 120 ml lipoaspirate. The cultured cells showed fibroblastic morphology which is characteristic for MSCs and were able to be purified from non-MSCs cells. This was confirmed by flowcytometry assay showing expression of CD105 and the absence of HLA-Class II, CD 45, CD 34, CD14, and CD19.Conclusions This study has shown that it was feasible to isolate messenchymal stem cell from human lipoaspirate. The procedure was practicable to be performed within a basic laboratory. (Med J Indones 2009; 18: 91-6)Keywords: mesenchymal stem cell, lipoaspirate, stem cell isolation technique

Immunogenicity characterization of mononucleated cells originated from umbillical cord blood Sardjono, Caroline T.; Setiawan, Melina; Suyatna, Frans D.; Japutri, Irvanyuni; Setiawan, Boenjamin; Sandra, Ferry
Medical Journal of Indonesia Vol 19, No 1 (2010): February
Publisher : Faculty of Medicine Universitas Indonesia

Aim Umbilical cord blood mononucleated (UCBMC) cells has been shown to be the stem cells originated from umbilical cord blood. To date, UCBMC has been introduced as an alternative source for stem cells used in autologous and allogeneic transplantations. Several clinical studies have demonstrated that UCBMCs required less stringent selection for HLA matches between donor and recipient with less cases of graft versus host reaction. In this study, UCBMCs are known to contain many stem cells, were characterized and compared to peripheral blood for their immunogenic profile.Methods To elucidate the potential of UCBMC alloreactivity, mixed lymphocyte reaction (MLR) assay was performed. The donor and effectors cells were HLA-typed using PCR method to determine their alloreactivity. Further, to distinguish the level of HLA class I and II expression flowcytometry was done using monoclonal antibodies against those molecules. All the analyse were carried out on UCBMCs and peripheral blood mononucleated cells (PBMCs).Results The result of MLR assay showed that there was less IFN-Î³ secretion detected in the co-cultured medium in the presence of UCBMCs compared to PBMCs counterpart, indicating less possible rejection of UCBMC. Further, we found that there were only 1-3 alleles of HLA match (out of 8 alleles) among the PBMCs and UCBMCs. By using flowcytometry assay, we could further demonstrate lower HLA Class I expression level with less amount of HLA Class II expressing cells in UCBMC compared to those in PBMCs.Conclusion These findings clearly demonstrate the low immunogenicity of UCBMCs, based on the low level of secreted IFN-Î³ in the MLR assay, low expression level of HLA Class I, and small population of HLA Class II expressing cells. The outcomes from this study would raise a better understanding in the usage of umbilical cord blood as an alternative source of stem cells for allogeneic transplantation. (Med J Indones 2010; 19:14-20)Keywords: umbilical cord blood, immunogenicity, stem cell

IFNÎ³ modulates human immunoglobulin receptor expression in lipoaspirate-derived mesenchymal stem cells Laksmitawati, Dian R.; Pawitan, Jeanne A.; Sadikin, Mohamad; Sardjono, Caroline T.; Utomo, Ahmad R.
Medical Journal of Indonesia Vol 23, No 3 (2014): August
Publisher : Faculty of Medicine Universitas Indonesia

Background:Â Mesenchymal stem cell (MSC) has been reported to have immunomodulator capacity against autoimmune diseases and to prevent allogenic tissue rejection. Many studies revealed that MSCâs inhibit T cell proliferation and induce immunosuppressive condition through the production of prostaglandins, and interleukin-10. In addition, MSC was reported to reduce circulating autoantibody in autoimmune patients following MSC transfusion. So far, there has been no report stating the presence of Fc receptors (receptors for immunoglobulin) on MSCs. The aim of this study was to reveal the expression of FcÎ³Rs in lipoaspirate-derived MSCs by measuring transcription of FcÎ³R mRNA and whether the expression can be modulated.Methods:Â Lipoaspirate-derived MSCs were cultured in suitable medium and confirmed to be MSCs according to the criteria published by International Society for Cellular Therapy. Total mRNA of MSCs was isolated, and detection of human FcÎ³RI, FcÎ³RIIA and FcÎ³RIIB mRNA was performed. Further, modulation of the expression was tested using heat aggregated gamma globulin (HAGG) and interferon (IFN)Î³.Results:Â FcÎ³Rs mRNA was detected in the first passage of MSCs. However, the expression was no longer present after more than 4 passages. Further, increased level of FcÎ³RI and FcÎ³RIIA mRNA expression was detected with the addition of IFNÎ³ in the culture. This preliminary finding opens a new insight for the understanding of interaction between MSCs and immunoglobulin G through FcÎ³Rs.Conclusion:Â Lipoaspirate-derived MSCs express FcÎ³Rs, and the expression is modulated by IFNÎ³.

The application of human umbilical cord blood mononuclear cells in the management of deep partial thickness burn Moenadjat, Yefta; Merlina, Maurin; Surjadi, Camy F.; Sardjono, Caroline T.; Kusnadi, Yuyus; Sandra, Ferry
Medical Journal of Indonesia Vol 22, No 2 (2013): May
Publisher : Faculty of Medicine Universitas Indonesia

Background: Wound healing in burn is a complex process and early complete wound closure still enfaces many problems. Application of stem cells is found to be the future method of wound healing. Among the available sources of allogenic stem cells, umbilical cord blood is quite easy to be obtained, has less ethical issue, and contain multipotent stem cells, which are characterized by low immunogenicity. The study aims to evaluate the potential of human umbilical cord blood mononuclear cells (hUCBMNCs) treatment in the management of deep partial thickness burns. Methods: Twenty patients with deep partial thickness burns were treated with topical application of 2 x 107 hUCBMNCs and silver sulfadiazine (SSD) cream on the comparable wound size in the other sites. The treatments were applied for six times in every two consecutive days. Wound surface area was measured with Visitrak® on day 0, 7, and 11. Pain intensity was evaluated using Wong Baker’s faces scale on each wound dressing change. Histology examination was performed in some samples of collected skin biopsy of the newly re-epithelialized area of hUCBMNCs and SSD-treated wound at the end of treatment. HLA typing is used to evaluate the issue of safety. Wilcoxon signed rank test was used to compare the rate of wound healing. Results: Sixteen patients of hUCBMNCs-treated showed a significant wound closure in faster than SSD-treated; measured on day 7 (p = 0.041) and day 11 (p = 0.021). Number of patients with reduced pain intensity, from approximately scale 3 to 1/0 on day 7 and 11, were higher in hUCBMNCs-treated compared to SSD-treated wound. In spite of the HLA-mismatch, no allergic reaction, rejection, and infection found on hUCBMNCs-treated wound suggested the safety of this therapy. Histology examination found the formation of dermal-epidermal junction and rete ridges equal to the normal skin on hUCBMNCs-treated wounds. Conclusion: hUCBMNCs are effective and safe to promote re-epithelialization in deep partial thickness burns. (Med J Indones. 2013;22:92-9)Keywords: Deep partial thickness burn, mononuclear cells, re-epithelialization, umbilical cord blood

Co-Authors Ahmad R. Utomo Boenjamin Setiawan Camy F. Surjadi Dian R. Laksmitawati Ferry Sandra Frans D. Suyatna Frisca Frisca Gwendy Aniko Irvanyuni Japutri Jeanne A. Pawitan Maurin Merlina Melina Setiawan Mohamad Sadikin Virgi Saputra Yefta Moenadjat Yuyus Kusnadi

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