MENSTRUAL BLOOD STEM CELLS (MenSCs)

T.N.JAYA GANESH,II-M.Sc BIOTECHNOLOGY,DEPT. OF BIOTECHNOLOGY,BHARATHIAR UNIVERSITY.

INTRODUCTION

A population of stromal stem cells derived from menstrual blood (MenSCs) are easily expandable to clinical relevance and express multipotent markers such as Oct-4, SSEA-4, and c-kit at the molecular and cellular level.

Moreover the multipotency of MenSCs by directionally differentiating MenSCs into chondrogenic, adipogenic, osteogenic, neurogenic, and cardiogenic cell lineages.

These studies demonstrate the plasticity of MenSCs for potential research in regenerative medicine.

SOURCES

Endometrium from the eutopic (inside the uterine cavity) ectopic (outside of the uterine cavity), from the menstrual blood and bone marrow are the origin of the putative

endometrial stem cells.

Isolation and differentiation of stromal cells

FACS

Antibodies for human cell surface antigens,CD9, CD29, CD34, CD38, CD44, CD45, CD49f, CD90, CD105, CD117, CD166, LIN, MHC I, and MHC II.

Also with c-kit and SSEA-4.

KARYOTYPE ANALYSIS

No chromosomal aberrations are expressed by these cells as determined by karyotype analysis at passage 12.

RT-PCR ANALYSIS

MenSCs express multipotent marker Oct-4 at the RNA level up to 12 passages as determined by Reverse Time-PCR L- ladder; W- water; ESC- embryonic stem cells; P12- passage 12

MenSCs stain positive for (a) c-kit, (c)SSEA-4 and (e) c-kit+ and SSEA-4 colocalization. Negative controls did not stain for c-kit or SSEA-4 (b, d, f). Scale bar: 20 µm.

MenSCs are easily differentiated into mesoderm tissue types. Staining with oil red O for fat vacuoles demonstrates differentiation into adipogenic tissue when induced (b) compared to noninduced controls (a). Staining with alcian blue for sulfated proteoglycans demonstrates differentiation into chondrogenic tissue types when induced (d) compared to noninduced MenSCs controls (c). Staining with alizarin red S (f) in induced cells shows calcium deposits when compared to the noninduced controls (e). Scale bar: 20 µm.

MenSCs differentiate into neural tissue. (a–c) Neural-induced MenSCs differentiate into oligodendroglial cells expressing markers O4 and GalC. Moreover, they can be differentiated into neuronal cells and neural progenitors shown by the expression of(d) Map-2, (e) Tub-3, and (f) Vimentin (F), respectively.

MenSCs express telomerase activity at passage 12. hESC, human embryonic stem cells; MEF, mouse embryonic fibroblasts.

EMBRYONIC STEM CELLS AND ADULT STEM CELLS

NOVEL PATHOGENESIS OF ENDOMETRIOSIS Endometriosis is a multifactorial disease, the pathogenesis includes retrograde

of endometrial cells, immunological insufficiency, metaplasiaand environmental hormone disruptor.

Endometrial stem cells were identified in the endometrioma cyst Some hypotheses suggest the theory of stem cell in the pathogenesis of endometriosis

Endometrium stem cells reside in the basalis layer can flux through the tubes and establish endometriotic lesions in the peritoneum.

The stimulatory factors consisting of genetic, immunological and inflammatory, angiogenesis factors promote the growth.

An alternative hypothesis explores the extrauterine stem/progenitor cells (for example, from reprogram of bone marrow MSCs and circulating EPCs) function in the pathogenesis of endometriosis.

Schematic representation of a decision tree based on six sources of MSCs and five different criteria related to their availability, isolation procedure and different properties

At class 7 clean room the cells are stored in two cryovials which are overwrapped to safeguard the cells during storage and a technique called “controlled-rate freezing” is used to prepare the cells for long-term storage. The overwrapped storage vials are housed in the vapor phase of liquid nitrogen for cryogenic preservation.

CONCLUSION

Have fast doubling time than HSC’s in 24-36hrs MenSCs have been tested only in very limited disease models. possess various regenerative properties under physiological and pathological

conditions. From a translational point of view, MenSCs appear to have practical and also

biological advantages over other stem cell sources. Easy collection of SC’s and can be preserved

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DISCUSSION